Objective To investigate the vascular endothelial growth factor (VEGF) expression level by chondrocytes isolated from patients with osteoarthritis (OA) in hip or femoral neck fracture (FNF) and explore the effect of synovial fluid from OA or FNF on secretion of VEGF. Methods The cartilage tissues were collected from 12 patients with OA in hip and 8 patients with FNF. Cartilage was stained with HIM and Safranin O/Fast Green (S/F) method. The damage of cartilage was evaluated using Mankin scores.Cathepsin B which was selected for cell dedifferentiation monitoring marker and VEGF level was detected in the supernatant fluid. The synovial fluid from OA, FNF and DMEM were respectively added to the culture medium to explore their effects on regulating VEGF. Results Cartilage the Mankin scores of OA group were higher than that of FNF group. Chondrocytes gradually lost their original spherical appearance, with Cathepsin B upregulated while VEGF downregulated. The OA synovial fluid can stimulate chongdrocytes to secrete more VEGF than the one from patients with FNF. However, chondrocytes gradually produced less VEGF after passaging. Conclusion Mankin scores had good correlation with chondrocytes' VEGF production in the early stage of primary culture. Chondrocytes showed quick dedifferentiation characteristics in vitro. OA synovial fluid showed abig ger capability in stimulating chondrocytes to express more VEGF, which might indicate that OA synovial fluid participated in the pathological process of OA.

Objective:To observe the bone marrow mesenchymal stem cell isolation,identification methods and protection mechanisms in the kidney of diabetic rats.Methods: Thirty-one rats were used to isolate bone marrow mesenchymal stem cells (BMSCs) by adherent culture method.Cell morphology was observed by phase contrast microscope.The surface molecular weight of cultured cells was detected by Real-time PCR.30 rats were induced by intraperitoneal injection of streptozotocin to establish diabetic rat model (n=15).And stem cell treatment group (n=15) according to the different treatment methods.Rats in the control group were treated with insulin combined with probucol.Stem cell treatment group was implanted with stem cells.Before and after treatment,fasting blood glucose,24 h urinary protein excretion,creatinine clearance rate and kidney weight were measured.PAS staining.And the renal tissue was detected by Western blot.Results: The primary bone marrow mesenchymal stem cells (MSCs) were cultured for 24 hours,and the cells were irregularly distributed in the culture flask.After 7 days of culture,the cells were regular,elongated,elliptical,and strong refraction,and the cells were fused with each other.The morphology of BMSCs was uniform and showed a long fusiform shape,CD44,CD31 and CD34 were observed by RT-PCR.The levels of fasting blood glucose,24 h urine protein excretion and creatinine clearance rate in the observation group were significantly higher than those in the control group (P<0.05).The results of PAS staining showed that the glomerular structure was clear,the morphology was regular,the arrangement rules of glomerular cells,the basement membrane and the encapsulation of the glomerular membrane were significantly higher than those of the control group Man′s capsule clear (P<0.05).In the control group,the glomerular volume increased,the mesangial area widened and the stroma increased significantly.The Bcl-2 protein level and the Bax and Caspase-3 protein levels were significantly decreased in the stem cell treatment group.The levels of Bcl-2 and Bax and Caspase-3 protein in the control group were significantly decreased (P<0.05).The ROS,MDA and SOD levels in oxidative stress group were lower than those in control group (P<0.05).The difference of ROS,MDA and SOD between the two groups was not statistically(P>0.05).Conclusion: It is suggested that BMSCs can be isolated from rat bone marrow mesenchymal stem cells by using adherent culture method and labeled with green fluorescent protein in vitro.Bone marrow mesenchymal stem cells can protect the kidney and inhibit the oxidative stress of diabetic kidney.

Objective:To investigate the effect of alprostadil combined with metformin in the treatment of diabetic nephropathy.Methods:50 cases of diabetic nephropathy patients were enrolled and then divided equally into the observation group and the control group. The patients in the control group were treated with conventional therapy and metformin, and the patients in the observation group were treated with alprostadil on the basis of the treatment of the control group. Compare the glycemic index, lipid index, renal function index, inflammatory response index, and oxidative stress response index of the two groups of patients before and after the 4-week treatment. The ratio of the number of effective cases (significant + effective) to the total number of cases, i.e., the total effective rate, was used to characterize the treatment effect.Results:The total effective rate in the observation group was higher than that in the control group (88.00% vs. 60.00%, P<0.05). After the 4-week treatment, no adverse effects occurred in either group. Compared with the control group, patients in the observation group had higher fasting blood glucose (FBG), glycosylated hemoglobin(HbA1c), mean blood glucose(MBG), blood glucose fluctuation rate(BGFR), standard deviation of blood glucose(SDBG), triglyceride(TC), total cholesterol(TG), high-density lipoprotein(HDL), low-density lipoprotein(LDL), serum creatinine(Scr), urinary microalbumin(UmALB), glomerular filtration rate(eGFR), albumin/creatinine ratio(ACR), cyclic adenosine monophosphate(cAMP), renin, protein kinase(PKA), epinephrine (E), angiotensin-converting enzyme inhibitor Ⅱ(ACEI Ⅱ), and norepinephrine(NE) were improved(all P<0.05). Conclusions:The effect of alprostadil combined with metformin in the treatment of diabetic nephropathy is accurate, safe, and reliable.

OBJECTIVETo analyze early hematopoietic response and long-term survival of very severe aplastic anemia (VSAA) patients with different absolute neutrophil counts (ANC) after frontline immnunosuppressive therapy (IST).

METHODSClinical data and outcome of 145 VSAA patients treated with rabbit antithymocyte globulin combined with cyclosporine were retrospectively analyzed. Hematopoietic responses to IST and long-term survival were statistically analyzed for VSAA patients in different ANC subgroups.

RESULTSPre-IST ANC=0.05×10(9)/L acted as the best cutoff level to predict IST response at 3, 6 months. For 145 VSAA patients, early death rate was 13.4% (11/82) vs 1.6% (1/63), respectively, in the ANC≤0.05×10(9)/L group and ANC>0.05×10(9)/L group (P<0.05). Hematopoietic response rates to IST was 22.0% vs 54.0% (P=0.000) at 3 months, 34.1% vs 63.5% (P=0.000) at 6 months; the overall five-year survival rate was only (62.5±5.4) % vs (91.4±3.7) % (P=0.000) and five-year event-free survival rate was (42.3±5.5) % vs (63.1±6.5) % (P=0.003), respectively, in the ANC≤0.05×10(9)/L group and ANC>0.05×10(9)/L group.

CONCLUSIONVSAA patients with extremely low ANC (≤0.05×10(9)/L) had high early death rate and with very low response rate to frontline IST and poor survival, so it is urgent to seek for the alternative frontline therapy that will bring faster and better outcome for these patients.

Anemia, Aplastic ; blood ; drug therapy ; Animals ; Antilymphocyte Serum ; therapeutic use ; Cyclosporine ; therapeutic use ; Disease-Free Survival ; Humans ; Immunosuppressive Agents ; therapeutic use ; Leukocyte Count ; Neutrophils ; cytology ; Rabbits ; Retrospective Studies ; Survival Rate ; Treatment Outcome

Objective:To investigate the nutritional risk, incidence of malnutrition, and intake of three major energy-supplying nutrients, analyze changes in their body composition and the possible influencing factors in patients with stomach neoplasms during perioperative period in order to provide a theoretical basis for the nutritional management of patients with stomach neoplasms during perioperative period.Methods:This was a cross-sectional study. A total of 105 patients who underwent gastric cancer radical surgery in the Gastrointestinal Department of Zhongshan Hospital Affiliated to Xiamen University from June 2021 to May 2023 were taken as the research subjects using fixed-point continuous sampling method. They were recruited for screening and assessment using Nutritional Risk Screening 2002 (NRS 2002) and Patient-Generated Subjective Global Assessment (PG-SGA). Nutrients intake during the perioperative period were investigated using the 24-h recall method and dietary diary method, etc. Body compositions were measured using the bioelectrical resistance method.Results:Among the 105 patients, there were 78 males and 27 females, with an average age of (61.5 ± 10.3) years. About 83.8% (88/105) gastric cancer patients were at nutritional risk and 82.9% (87/105) were malnourished. The preoperative and postoperative energy intake were (1 646.1 ± 321.5) and (1 317.2 ± 365.8) kcal (1 kcal=4.184 kJ), respectively, which were significantly lower than the target amount of (1 896.7 ± 262.9) kcal, the difference was statistically significant ( t=6.23, 8.29, both P<0.05).The preoperative body mass, muscle mass, skeletal muscle, fat mass, and skeletal muscle index were (51.5 ± 9.6), (40.8 ± 6.0), (23.6 ± 4.0), (8.3 ± 4.9) kg, and 6.7 ± 0.8 respectively, while the postoperative values were (50.0 ± 9.1), (39.8 ± 6.0), (22.8 ± 3.6), (7.8 ± 5.2) kg, and 6.5 ± 0.8 respectively, with statistically significant differences between the two groups ( t values were 2.89-10.61, all P<0.05). Logistic multivariate regression analysis showed that the operation time ( OR=3.984, 95% CI 1.433-11.080, P<0.05) and energy satisfaction ( OR=0.053, 95% CI 0.005-0.610, P<0.05) were independent influencing factors for the degree of skeletal muscle loss. Conclusions:During perioperative period, the gastric cancer patients had poor nutritional status with insufficient nutrient intake and accelerated loss of body muscle and fat. Therefore, it was necessary to conduct a comprehensive nutritional evaluation for patients with stomach neoplasms during perioperative period in time and take steps to promote recovery by providing individualized nutritional therapy.

BACKGROUND:The study of the physical properties of scaffolds has always been a hot topic in the field of tissue engineering research.However,for vascular stimulating scaffolds,in addition to meeting the basic performance of the scaffold,other methods are also needed to promote the regeneration of blood vessels within the scaffold,in order to achieve the ultimate goal of repairing bone tissue. OBJECTIVE:A visualization analysis was carried out on the literature published in and outside China on scaffold stimulation for bone tissue engineering,to explore the research hotspots and research status in this field,and to provide a reference for subsequent studies. METHODS:Using the CNKI database and Web of Science core database as retrieval databases,the relevant literature on vascular scaffolds for bone tissue engineering was retrieved.The literature that did not conform to the research object was removed.The obtained data were imported into CiteSpace 6.1.R2 software.Visualization analysis was performed on authors,national institutions,and keywords in the research field. RESULTS AND CONCLUSION:(1)China,the United States,and Germany were the top three countries with the most articles on scaffold stimulation for bone tissue engineering.(2)The top 3 institutions in the CNKI database were Southern Medical University,Huazhong University of Science and Technology,and Donghua University.In the core database of Web of Science,Shanghai Jiao Tong University,Sichuan University and Chinese Academy of Sciences ranked the top 3 in terms of the number of institutional publications.(3)The top 3 keywords in the CNKI database were"tissue engineering,vascularization,angiogenesis".The top 3 keywords in the Web of Science core database were"mesenchymal stem cell,scaffold,vascularization".(4)Through the analysis of co-citation and highly cited references,the main concerns were as follows:vascularization strategies:scaffold design,angiogenic factor delivery,in vitro co-culture,and in vivo pre-vascularization.Technology:3D printing,electrospinning,vascular transplantation,vascular fusion.Mechanisms:immune regulation and macrophages,drug/growth factor delivery,the relationship between endothelial cells and osteoblasts,the paracrine relationship between bone cells and endothelial cells,signaling molecular pathways,angiogenesis,and anti-angiogenesis molecules.(5)The researches concerning vascular stimulating scaffolds in bone tissue engineering in and outside China attach great importance to the application of stem cells and 3D printing technology.Current research focuses on biological 3D printing technology,scaffold modification methods,and the development and application of intelligent biomaterials based on bone repair mechanisms.

Objective:To solve the difficult problem of experimental teaching of MRI equipment by independently designing a virtual simulation teaching system for performance testing of medical magnetic resonance imaging (MRI) equipment.Methods:A total of 202 students of Batch 2016 majoring in 4-year medical imaging technology and 5-year medical imaging of Binzhou Medical University. According to the teaching requirements of MRI equipment in Medical Imaging Equipment Science, a 3D experimental simulation model and experimental scenes were established based on Unity3D engine by using Unity3D, 3D Studio Max, Maya and Visual Studio technology to design experimental learning content and assessment content, develop the teaching system software and perform Web-based online learning. Then, the satisfaction survey on experimental learning was conducted, the statistics of the experimental results of the teaching system on the national virtual simulation experiment teaching platform was collected and the learning effects were evaluated in multiple dimensions. Results:The virtual simulation teaching system was a comprehensive experiment with 12 knowledge points, 12 experimental items and a total of 81 interactive contents and steps. The overall satisfaction rate of 202 students in our school was 96.82%(2 347/2 424). Among the 499 subjects who participated in the learning of the teaching system on the national virtual simulation experiment teaching platform and submitted the experimental report, the average score of the experiment was 78.07 points (the full score of online learning assessment was 90 points), and the overall passing rate was 96.79%(483/499). The average learning time of the 18 students who got 90 points was 54 min.Conclusion:This virtual simulation teaching system possesses comprehensive experimental contents with high fidelity of experimental scenes and strong interactivity of experimental operation, prospecting great learning effects and promotion value.

Clinical data of 19 patients with congenital pyruvate kinase deficiency were analyzed. Insufficient pyruvate kinase confirmed the diagnosis. Laboratory parameters of hemolysis were summarized. In cases of neonatal hyperbilirubinemia and unexplained hemolytic anemia, pyruvate kinase activity and next generation sequencing test may help the early diagnosis.

Objective: To evaluate the impact of the targeted next-generation sequencing (NGS) assay for difficult congenital anemias. Methods: Blood Disease Hospital Anemia Panel 2014 (BDHAP-2014) including 217 known genes of congenital anemias was developed. NGS and parental verification were performed for patients who were suspected diagnosed with congenital anaemia from August 2014 to July 2017. Results: A total of 46 patients were enrolled in this study, the clinical suspection were 11 cases Fanconi anemia (FA), 8 cases congenital dyserythropoietic anemia (CDA), 6 cases congenital sideroblast anemia (CSA), 12 cases congenital hemolytic anemia (CHA), 1 case dyskeratosis congenital (DC), 4 cases iron-refractory iron deficiency anemia and 4 cases unexplained cytopenia (Uc), respectively. 28 (60.9%) of 46 patients became confirmed cases after targeted NGS, corresponding to 44 mutations of which 33 were new. 26(56.5%) patients with results of the assay matching to clinical suspection, including FA (5/11, 45.5%), CSA (6/6, 100.0%), CDA (3/8, 37.5%) and CHA (12/12, 100.0%). 2 (4.3%) cases not matching to clinical suspection, including dyskeratosis congenital (DC) was made in 1(2.2%) patients with suspected FA and familial hemophagocytic lymphohistiocytosis (FHL) was made in 1(2.2%) patients with suspected unexplained cytopenia (Uc). In 12 CHA patients, the hemolytic type was further clarified by the NGS. The remaining 18 cases were not clearly diagnosed. Conclusion Targeted NGS assay is of major impact on congenital anemias. The assay should be used routinely in congenital anemias.

Objective: To reveal the genetic characteristics of erythrocyte membrane protein in hereditary spherocytosis (HS) in China. Methods: Next-generation sequencing technology was used to detect mutations in genes of erythrocyte membrane proteins in 51 clinically diagnosed HS patients. The relationship between gene mutations and clinical phenotypes was analyzed. Results: Mutations in erythrocyte membrane protein genes were detected in 37 patients, including 17 with ANK1 mutations (17/37, 45.9%), 14 with SPTB mutations (14/37, 37.8%), and 5 with SLC4A1 mutations (5/37, 13.5%). One patient carried both heterozygous ANK1 mutation and SPTB mutation (1/37, 2.7%). SPTA1 and EPB42 mutation was not fou nd in any patient. Nonsense mutations (36.8%) and missense mutations (31.6%) were most common. Of the 38 mutations detected, 34 were novel mutations and have not been reported elsewhere (89.5%). Sixteen HS patients underwent parental genetic validation, 6 patients (37.5%) inherited gene mutation from parents and 10 (62.5%) were de novo. The peripheral blood cell parameters of HS patients were not related to the mutant genes and gene mutation types. However, it seems that HS patients with mild clinical status are prone to carry SPTB mutations while more patients with severe clinical status have ANK1 mutations. Conclusions ANK1 and SPTB are the most common mutant genes in Chinese HS patients, mainly with missense mutations and nonsense mutations. There was no significant correlation between the mutation of HS related genes and the severity of HS.