Objective To explore the age-related biological properties of bone-derived mesenchymal stem cells(BMSCs)from mice of different age groups and their osteogenic differentiation induced by bone morphogenetic protein 2(BMP2).Methods Eight C57BL/6J mice were divided into a young group(4 weeks old,weighing 10～15 g,n=4)and an old group(12 months old,weighing 20～25 g,n=4),with half male and half female.MSCs were extracted from the whole bones of the 2 groups of mice.After the obtained cells were identified with flow cytometry for the surface markers,β-galactosidase staining was employed to compare the senescence level of BMSCs,MTT and EdU incorporation assays were conducted to compare the proliferation and self-renewal abilities of between the 2 groups.Western blotting was employed to analyze the expression of CyclinD1 and P21 in BMSCs.Then ALP staining,Alizarin Red staining and RT-qPCR were used to evaluate the osteogenic differentiation ability of the cells.RNA sequencing was performed to compare the differential gene expression in BMP2-induced BMSCs.Lastly,the sequencing results were re-confirmed by using flow cytometry.Results Flow cytometry showed that the sorted and cultured mouse BMSCs met the criteria for MSCs.The results of β-galactosidase staining indicated that the senescence level of BMSCs in the old group was significantly higher than that in the young group(P＜0.05).MTT and EdU doping experiments revealed that the cell viability and proliferation ability of BMSCs were significantly lower in the old group than the young group(P＜0.05).Western blotting displayed that the expression level of cell cycle protein CyclinD1 was lower,whereas that of cell cycle inhibitory factor P21 was significantly higher in the BMSCs from the old group than the cells from the young group(P＜0.05).ALP/Alizarin Red staining and RT-qPCR demonstrated that the BMSCs from the young group had stronger osteogenic differentiation capacity after BMP2 treatment when compared the cells of the old group(P＜0.05).RNA sequencing results displayed that the changing profile of CD51 expression was in opposite trends in the young and old BMSCs after BMP2 treatment.Finally,flow cytometry revealed that the percentage of CD51+cells within the CD45-cells was significantly higher in the young group than the old group.Conclusion The decrease in the percentage of CD51+cells among CD45-cells in aged BMSCs is closely associated with their decreased responsiveness to BMP2-induced osteogenic differentiation.

Mice ; Animals ; Rett Syndrome ; Longevity ; Methyl-CpG-Binding Protein 2 ; Disease Models, Animal ; Phenotype

Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines

Objective To investigate the normal reference values of abdominal aorta and its branches in Southwest China, so as to provide basis for clinical diagnosis and treatment of abdominal aortic aneurysm. Methods The length, diameter and angle of the abdominal aorta and its branches were measured in 20 cadavers. At the same time, 300 cases without obvious pathological changes in four urban populations were selected for abdominal CT angiography (CTA) to measure the length, diameter and angle of abdominal aorta and its branches. Results Display rate of abdominal aorta was 100%,the average length of abdominal aorta was (12.96±0.81) cm, the diameter of abdominal aorta was (1.93±0.28) cm, (1.55±0.23) cm, (1.61±0.19) cm in renal artery plane (Ⅰ), inferior mesenteric artery plane (Ⅱ) and lower edge plane of lumbar vertebrae (Ⅲ), and the average value on image was (2.17 ± 0.42) cm, (1.81 ± 0.40) cm, (1.99 ± 0.53) cm; Display rate of common iliac artery was 100%, the angle between left and right common iliac artery was (59.80± 4.66) ° and the diameter of the end was (1.16±0.33) cm and (1.12±0.11) cm; Display rate of celiac trunk on cadaver was 100%, the average length was (1.61±0.27) cm, the average diameter on cadaver was (0.78±0.71) cm, the image was (0.88± 0.31) cm; Display rate of renal artery was 100%, the average length of the left renal artery was (2.58 ± 0.50) cm, that of the right was (4.26±0.65) cm, the initial average diameter of the left renal artery was (0.55±0.24) cm, and the right renal artery was (0.62±0.20) cm; Display rate of the superior mesenteric artery was (100%), that of the on cadaver was (4.56 ± 0.29) cm, that of the image was (4.93 ± 0.84) cm, and that of the angle between the abdominal aorta was (38.05±5.99)°; Display rate of the inferior mesenteric artery was (100%),the average length of the inferior mesenteric artery was (6.57±0.79) cm on the cadaver, the image was (6.70±0.76) cm, and that of the angle between the abdominal aorta was (73.79 ± 9.62) ° and the initial diameter was (0.48 ± 0.29) cm. Conclusion The normal reference values of abdominal aorta and its visceral branches in Southwest China population were measured. CTA has a special advantage in showing the lesions of abdominal aorta and its branches in lumen, extraluminal and vascular wall, which can provide anatomical data of patients before operation.

Objective To investigate the clinical value of amplitude integrated electroencephalogram on early diagnosis and prognosis evaluation of brain injury caused by neonatal asphyxia.Methods A total of 34 full-term asphyxiated neonates(asphyxia group)hospitalized in NICU of our hospital from January 2015 to September 2015 were selected;meanwhile,34 full-term healthy infants(control group)of the same term were selected.All cases were monitored for the activities of aEEG background,sleep-awakening cycle(SWC)and epileptic activity(SA)within 6 hours after birth.Meanwhile,the relationships between various indexes and asphyxia degree and brain injury were analyzed.Results The electroencephalogram of the asphyxia group was 52.9％and the rate of SWC was 58.8％,which were lower than those of the control group,and the difference had statistic significance(P<0.05).Meanwhile,neonates with epileptic activity in asphyxia group accounted for 11.8％,which was higher than that of control group significantly(P<0.05).Conclusion The AEEG changes of neonates at early period after birth are closely related to perinatal asphyxia and brain injury after asphyxia.The application of amplitude integrated electroencephalogram has an important significance on early diagnosis of neonatal asphyxia.

Objective To explore the experience of diagnosis and surgical treatment of carotid body tumor.Methods A retrospective analysis between November 2008 and November 2015 was proceeded,the clinical data of surgical treatment for 81 patients with carotid body tumor was collected,to analyze data by SPSS19.0,and summarize the diagnosis of carotid body tumor,choice of operation methods and curative effect and complications prevention.Results Seventy-four cases underwent surgery treatment:tumors of 52 cases were simply stripped,tumors of 13 cases were resected combined with ligation of external carotid artery.Tumors of 7 cases were resected with internal and external carotid artery ligation,3 cases of whom underwent artificial blood vessel internal carotid artery end to end anastomosis.Postoperative death in 1 case of acute myocardial infarction,complicated with cerebral infarction in 2 cases,6 cases of injury of cranial nerve relieved after symptomatic treatment.No hemiplegia,aphasia and other serious complications.Tumor size and the surgery time correlation analysis:the correlation coefficient was 0.226,no significant correlation.Conclusions CTA is the most commonly used method of preoperative examination.Surgical resection is an effective method in treatment of carotid body tumor.Prevention injury of carotid artery cr internal carotid or common carotid artery and their reconstruction is the key to a successful operation.Sufficient preoperative assessment,select the appropriate operation method,intraoperative careful performance can ensure the cerebral perfusion,is the key to prevent and reduce the complications.

Objective Tocheck and evaluate on clinical practice teaching files for providing a foundation of improved clinical teaching levels with enhanced establishment of practice teaching files. Methods The practice teaching files of eighteen clinical departments in a military medical university were selected as the subject and were evenly divided into teaching plans and teaching records as well as teach-ing summary three parts and 15 items. Each item had 2 scores in which quantity had one score and quality has another one. Evaluation results were converted into 100 points system. Results Among practice teach-ing files of eighteen clinical departments, 4 scored more than 90, 8 scored between 80 to 89, 3 scored between 70 to 79, 3 scored between 60 to 69. Three advanced departments are obstetrics and gynecology, auxiliary department and emergency department. There were no significant differences in the scores of major surgery and internal medicine. Compared with that of other departments in the hospital, the specialty of hospital had low scores. It showed that most of the clinical practice of the file information was relatively sound with complete records and some departments had missing items or incomplete and non-standard doc-umentation records. Conclusion Qualitative study on practice teaching files can reflect clinical practice teaching quality. And it is meaningful for us to enhance teaching files construction and to improve clinical practice teaching levels.

Objective To investigate the diagnosis and treatment of visceral artery aneurysms (VAAs).Methods From June 2008 to December 2013,20 patients (11 male and 9 female) with visceral artery aneurysms were treated in our hospital.The clinical data and treatment outcomes were retrospectively analyzed.Results There were 20 patients,including 11 males and 9 females.The site of aneurysmal disease was splenic artery in 12 cases,mesenteric artery in 2 cases,hepatic artery in 2 cases,renal artery in 2 cases,celiac artery in 1 case and gastroduodenal artery 1 case.10 patients were treated by open surgery,and other 10 patients treated by endovascular.1 patient died,and others were discharged.All patients were followed up 28.5 months (ranged from 10 to 76 months).Two patients died due to cirrhosis of the liver,and the remaining patients were alive,no treatment-related complications,no serious complications,such as aneurysm recurrence occurs.Conclusions Both traditional surgical and endovascular treatment of visceral artery aneurysms may obtain a satisfactory outcome.

Objective To construct a recombinant bacillus Calmette-Guérin(BCG) vaccines based on different tandem repeats of MUC1 and GM-CSF, rBCG-MVNTR1/4/8-CSF, and to observe the ability of three recombinant BCG vaccines in the inhibition of breast cancer. Methods After MUC1 variable-number tandem repeats (MVNTR1/4/8) were cloned in a stepwise manner, the E. coli-Mycobacteria shuttle expression vector pDE22-MVNTR1/4/8-CSF were constructed by fusing MVNTR1/4/8 and GM-CSF, and then used to transform competent BCG by electroporation after identification by restriction endonuclease digestion analysis and DNA sequencing. A novel breast cancer vaccines, rBCG-MVNTR1/4/8-CSF was constructed. The expression of fused MVNTR1/4/8-CSF protiens was analyzed by SDS-PAGE and Western blot. The ability of rBCG vaccines inhibiting the growth of breast cancer was observed in hu-PBL-SCID mice. The specific T cell responses in mice were assessed by immunohistochemistry. Results The expression of recombinant MVNTR1/4/8-CSF fusion proteins were detected by SDS-PAGE and Western Blot in rBCG-MVNTR1/4/8-CSF vaccines, respectively. Tumor incidence in mice prophylactic immunized with rBCG-MVNTR4-CSF (37.5%) or rBCG-MVNTR8-CSF (25%) significantly decreased compared with PBS and BCG-pDE22 control ( 100% ) at 42 days after tumor implantation ( P ＜ 0. 05 ). MCF-7 tumor growth inhibition in rBCG- MVNTR4/8-CSF-immunized mice was more significant than that in controls ( P ＜0. 01 ).The inhibition effect of three rBCG vaccines on breast rumor growth appeared to rise with increase of numbers of the tandem repeats of MUC1. Survival rate was 75% of mice in the rBCG-MVNTR4-CSF-treated group and 87. 5% of mice in the rBCG-MVNTR8-CSF-treated groups at 70 days after tumor implantation; however,survival rate was only 12. 5% in control group( P ＜0. 05). The CD4-positive and CD8-positive lymphocytes were detected only in rBCG-MVNTR4/8-CSF-immunized mice. Conclusions rBCG-MVNTR4/8-CSF immunization inhibits breast cancer growth in mice.

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.