BACKGROUND:Primary cortical neurons and microglial cells play a crucial role in exploring cell therapies for neurological disorders,and most of the current methods for obtaining the two types of cells are cumbersome and require separate extraction.It is therefore crucial to find a convenient and rapid method to extract both types of cells simultaneously. OBJECTIVE:To explore a novel method for simultaneous extraction of primary cortical neurons and microglial cells. METHODS:Newborn suckling SD rats were taken within 24 hours.The brain was removed and placed in a dish with DMEM,and the pia mater was removed for later use.Primary neurons were extracted from the same brain tissue,and then the remaining brain tissue was used to extract microglial cells.The whole process was performed on ice.Extraction and culture steps of primary cortical neurons:The cerebral cortex was taken 2.0-3.0 mm with forceps,and the tissue was digested with papain for 20 minutes.After aborting digestion,the blown tissue presented an adherent tissue suspension.The supernatant cell suspension was obtained,filtered,and dispensed into 15 mL centrifuge tubes.After centrifugation and re-suspension,the cells were inoculated onto 6-well plate crawls coated with L-polylysine.Neuronal morphology was observed at 1-day intervals,and staining could be performed for identification using immunofluorescence staining of MAP2 and β-Tubulin by day 7.Microglia extraction and culture steps:The remaining brain tissue at 8-10 mm thick was subjected to microglial cell extraction,digested by trypsin for 20 minutes.After digestion was stopped,the tissue was blown to a homogenate,and then the homogenate was transferred to the culture bottle for culture.On day 14,the culture flasks were sealed and subjected to constant temperature horizontal shaking for 2 hours.Microglial cells were shed in the supernatant.Purified microglial cells were taken and continued to be cultured for 3 days for identification by Iba1 immunofluorescence staining. RESULTS AND CONCLUSION:(1)After 24 hours of culture,the neurons were adherent to the wall,the cytosol was enlarged,and some neurons developed synapses.After 3 and 5 days of culture,the cytosol was further enlarged,and most of the neurons were in the form of synapses,and some neurons were growing in clusters.On day 7,neuronal synapses were prolonged and thickened,and they were connected with each other to form a network.The neurons were identified by β-Tubulin and MAP2 immunofluorescence staining.(2)The cells grew close to the wall on day 1 of culture.On days 3,5,and 7,the density of microglial cells was small,and the cell morphology was bright oval or round,but the cells basically grew in clumps on the upper layer of other cells.On day 10,the density of microglial cells increased significantly.On day 14,microglial cells grew in dense clumps on the upper layer of other cells,and then they could be isolated and purified.The isolated and purified cells were taken and re-cultured to day 3 and identified as microglial cells by Iba1 immunofluorescence;their purity was greater than 95%.(3)The results show that primary cortical neurons and microglial cells obtained by this method after extraction and culture are of high purity,good morphology,and high viability.

OBJECTIVE: To summarize the clinical characteristics of foot and ankle deformities combined with knee and lower limb deformities and evaluate the advantages, clinical outcomes, and considerations of QIN Sihe's surgical strategy for treating such complex deformities. METHODS: Between January 2022 and December 2024, 32 patients with foot and ankle deformities combined with knee and lower limb deformities were enrolled. The cohort included 23 males and 9 females, aged 10-67 years (mean, 41.1 years). The main etiologies included post-polio sequelae (20 cases) and congenital limb deformities (3 cases). Deformities were categorized as follows: equinovarus foot (12 cases), equinus foot (2 cases), equinovalgus foot (3 cases), equinus foot with swan-neck deformity (2 cases), calcaneus foot (5 cases), foot valgus (2 cases), knee flexion deformity (14 cases), genu recurvatum (4 cases), genu varum (3 cases), genu valgum (3 cases), lower limb shortening (3 cases), and lower limb external rotation (6 cases). QIN Sihe's surgical strategies included osteotomies, tendon releases, and tendon transfers for deformity correction, followed by external fixation for residual deformity adjustment and stabilization. Outcomes were assessed using QIN Sihe's Postoperative Evaluation Criteria for Lower Limb (Foot and Ankle) Deformity Correction and Functional Reconstruction. RESULTS: All patients were followed up 8-32 months (mean, 16.5 months). Complications included pin tract infection (1 case, 1 site), ankle pain (2 cases), delayed healing at the proximal tibial osteotomy site (1 case), and anterior talar dislocation (1 case). At last follow-up, insufficient correction of foot deformity was observed in 1 case; both knee and lower limb deformities were corrected, with only mild recurrence of knee flexion deformity in 1 case. The foot/ankle and knee joint function improved. Based on QIN Sihe's Postoperative Evaluation Criteria for Lower Limb (Foot and Ankle) Deformity Correction and Functional Reconstruction, outcomes were rated as excellent in 30 cases and good in 2 cases, with an excellent-good rate of 100%. CONCLUSION Foot and ankle deformities combined with knee and lower limb deformities are complex, QIN Sihe's surgical strategy can achieve satisfactory clinical outcomes for simultaneous correction.
Humans ; Male ; Female ; Adult ; Middle Aged ; Child ; Adolescent ; Aged ; Treatment Outcome ; Young Adult ; Plastic Surgery Procedures/methods* ; Lower Extremity Deformities, Congenital/surgery* ; Osteotomy/methods* ; Foot Deformities, Congenital/surgery* ; Ankle Joint/surgery* ; Knee Joint/surgery* ; Foot Deformities/surgery*

Objective:To compare the efficacy and safety of azacitidine combined with venetoclax or CAG regimen in the treatment of newly treated elderly patients with acute myeloid leukemia (AML).Methods:A retrospective cohort study was conducted. The clinical data of 34 newly treated elderly patients with AML treated in the Fifth People's Hospital of Datong from May 2018 to August 2023 were retrospectively analyzed. According to the treatment regimen, all patients were divided into venetoclax group (azacitidine + venetoclax, 17 cases) and CAG group (azacitidine + CAG regimen, 17 cases). The clinicopathological characteristics, efficacy, adverse reactions and survival of the both groups were compared.Results:There were no statistically significant differences in the clinical data of both groups (all P > 0.05). The complete remission (CR) rate and the objective response rate (ORR) in venetoclax group were higher than those in CAG group [CR: 70.6%(12/17) vs. 47.1% (8/17); ORR: 82.4% (14/17) vs. 64.7% (11/17)],while the differences in CR and ORR were not statistically significant (χ 2 = 2.00, P = 0.163; χ 2 = 2.00, P = 0.244). The follow-up time[ M ( Q1, Q3)] was 25.4 months (7.2 months, 60.3 months). At the end of follow-up, 19 of 34 patients survived (13 cases in venetoclax group and 6 cases in CAG group); 15 died (4 cases in venetoclax group and 11 cases in CAG group). The median overall survival (OS) time was 14.22 months (95% CI: 8.2-60.3 months) and 10.56 months (95% CI: 7.2-50.2 months), respectively in venetoclax group and CAG group;the median progression-free survival (PFS) time was 9.97 months (95% CI: 5.4-40.5 months) and 6.82 months (95% CI: 5.0-36.2 months), respectively, and there were no statistically significant differences in OS and PFS between the two groups (all P > 0.05). Grade 3-4 hematological adverse reactions occurred in 16 and 14 patients in venetoclax group and CAG group, respectively. There were no significant differences in granulocyte deficiency time, platelet deficiency time, infection and bleeding incidence between the two groups (all P > 0.05). Conclusions:Azacitidine combined with venetoclax or CAG regimen have better clinical efficacy and safety for newly treated elderly patients with AML.

Objective To construct models of viral transfection and hypoxia/reoxygenation induced cellular injury in adult mouse cardiomyocytes(AMCMs)isolated using a non-Langendorff method.Methods AMCMs were isolated,extracted,sedimented,and plated using a non-Langendorff method.The morphology and survival rate of the isolated cells were evaluated 2,24,48 and 72 h after plating,and their integrity was observed by immunofluorescence staining for α-actinin.The isolated AMCMs were infected with adenoviruses carrying an RFP-expressing vector and fluorescence images were obtained at 36 and 48 h post-infection and used to calculate transfection efficiency.The cells were cultured under hypoxic conditions for 45 min,reoxygenated for 24 h,and then stained with propidium iodide(PI)to verify establishment of the hypoxia/reoxygenation injury model.Results The survival rates of AMCMs at 2,24 and 48 h after plating were comparable,but survival was significantly reduced at 72 h.The integrity of the AMCMs was good and＞80%of the cells were transfected with adenovirus at 48 h.After hypoxia/reoxygenation treatment,42%of cells were stained by PI,suggesting successful establishment of the AMCM injury model.Conclusions In this study,we developed a non-Langendorff method for the fast and easy isolation of AMCMs with high cell viability.The isolated cells can be efficiently infected with adenovirus and respond to hypoxia/reoxygenation injury.These findings provide a systematic method for isolating AMCMs and for applying gene modification and hypoxia/reoxygenation injury in these cells.

Objective The objective of this study is to examine the expression level of the S100A7A protein in both gastric cancer tissues and cells,as well as to evaluate its impact on the malignant phenotype of gastric cancer(GC)cells.Methods Immunohistochemical assay was used to detect the expression characteristics of S100A7A in 21 gastric cancer tissues and their corresponding paracancerous tissues,as well as to investigate its correlation with gastric cancer clinicopathological factors.Gastric cancer cells were genetically modified to overex-press S100A7A through plasmid transfection.Subsequently,the impact of S100A7A on the proliferation,migra-tion,and invasion capacities of gastric cancer cells was assessed using cell proliferation assays(EdU assay and plate cloning assay)as well as cell migration and invasion assays(Transwell assay and scratch assay).Results The expression of S100A7A protein was higher in GC tissues than in paracancerous tissues;Overexpression of S100A7A may increase gastric cancer cell proliferation,migration,and invasion.Conclusion S100A7A is a possible oncogene in GC and is predicted to serve as a new diagnostic and therapeutic target for the disease.

In the field of health,with the innovation drive as the core,the new quality of productivity has successfully gets rid of the shackles of traditional health service delivery mode and industrial development trajectory,showing high-tech,high-efficiency and high-quality characteristics,which is in line with the development concept advocated in the new development stage,representing an advanced productivity quality.In the field of health economics,the new-quality productivity will have a decisive impact on the supply and demand of health services,the allocation and optimization of health resources,health policies and intervention measurement,health system performance and health equity.Meanwhile,the new quality productivity has a profound impact on health economic evaluation and technology evaluation,and even changes the health economic research paradigm.It is undoubtedly an important responsibility and challenge for the health economy research field to give timely and in-depth theoretical discussion on the new quality productivity and actively promote its practical application and development.It requires us to keep up with the pace of the times at the theoretical level and dig deep into the inherent law and value of the new-quality productivity.At the same time,in practice,it is committed to guiding and promoting the effective transformation and implementation of new-quality productivity in the field of health,so as to facilitate the elevation of health economic research and practice to a new qualitative level.

Developing new quality productive forces is an inherent requirement and important focus for promoting high-quality development.It revolves around the logical idea of"what-why-how to do",explains the connotation and characteristics of new quality productivity,proposes an understanding and recognition of the concept and characteristics of new quality productivity in internal auditing of public hospitals,and analyzes the theoretical logic and practical significance of developing new quality productivity in internal auditing of public hospitals.It further explores the specific path of developing new quality productivity in internal auditing of public hospitals,in order to provide reference ideas for the development of new quality productivity in internal auditing and empowering the high-quality development of internal auditing.

In the field of health,with the innovation drive as the core,the new quality of productivity has successfully gets rid of the shackles of traditional health service delivery mode and industrial development trajectory,showing high-tech,high-efficiency and high-quality characteristics,which is in line with the development concept advocated in the new development stage,representing an advanced productivity quality.In the field of health economics,the new-quality productivity will have a decisive impact on the supply and demand of health services,the allocation and optimization of health resources,health policies and intervention measurement,health system performance and health equity.Meanwhile,the new quality productivity has a profound impact on health economic evaluation and technology evaluation,and even changes the health economic research paradigm.It is undoubtedly an important responsibility and challenge for the health economy research field to give timely and in-depth theoretical discussion on the new quality productivity and actively promote its practical application and development.It requires us to keep up with the pace of the times at the theoretical level and dig deep into the inherent law and value of the new-quality productivity.At the same time,in practice,it is committed to guiding and promoting the effective transformation and implementation of new-quality productivity in the field of health,so as to facilitate the elevation of health economic research and practice to a new qualitative level.

Developing new quality productive forces is an inherent requirement and important focus for promoting high-quality development.It revolves around the logical idea of"what-why-how to do",explains the connotation and characteristics of new quality productivity,proposes an understanding and recognition of the concept and characteristics of new quality productivity in internal auditing of public hospitals,and analyzes the theoretical logic and practical significance of developing new quality productivity in internal auditing of public hospitals.It further explores the specific path of developing new quality productivity in internal auditing of public hospitals,in order to provide reference ideas for the development of new quality productivity in internal auditing and empowering the high-quality development of internal auditing.

In the field of health,with the innovation drive as the core,the new quality of productivity has successfully gets rid of the shackles of traditional health service delivery mode and industrial development trajectory,showing high-tech,high-efficiency and high-quality characteristics,which is in line with the development concept advocated in the new development stage,representing an advanced productivity quality.In the field of health economics,the new-quality productivity will have a decisive impact on the supply and demand of health services,the allocation and optimization of health resources,health policies and intervention measurement,health system performance and health equity.Meanwhile,the new quality productivity has a profound impact on health economic evaluation and technology evaluation,and even changes the health economic research paradigm.It is undoubtedly an important responsibility and challenge for the health economy research field to give timely and in-depth theoretical discussion on the new quality productivity and actively promote its practical application and development.It requires us to keep up with the pace of the times at the theoretical level and dig deep into the inherent law and value of the new-quality productivity.At the same time,in practice,it is committed to guiding and promoting the effective transformation and implementation of new-quality productivity in the field of health,so as to facilitate the elevation of health economic research and practice to a new qualitative level.