BACKGROUND:Currently, there is no uniform, standardized approach to isolate, purify and proliferate bone marrow mesenchymal stem cells. Chlormethylbenzamido-1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (CM-Dil) is a stable, reliable, high marking and simple marker.
OBJECTIVE:To develop the methods for isolation, culture and identification of rat bone marrow mesenchymal stem cells in vitro.
METHODS:Two male Sprague-Dawley rats, weighing 50-100 g were taken to col ect the bilateral femur and tibia bone marrow under sterile conditions, and then, primary bone marrow mesenchymal stem cells were isolated and cultured using bone marrow adherent separation and density gradient centrifugation. cells were amplified and purified through timely and repeated passage, and labeled at the third generation with fluorescent dyes CM-Dil in vitro as a source of donor cells.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells were cultured successful y in vitro using bone marrow adherent separation and density gradient centrifugation separation methods, but the former was superior to the latter in the number of cultured cells significantly, while the two methods were not different significantly in terms of cellviability and proliferation. Flow cytometry results showed that the positive rates of cultured cells were 17.5%for CD34, 97.9%for CD44, and 91%for CD90. CM-Dil can label bone marrow mesenchymal stem cells successful y, which is a stable, reliable, high marking and simple marker.

OBJECTIVE: Computer has become an important tool in medical works. However, application ability of clinical physicians is imbalanced with the fast development of information technology (IT) system. Therefore, it is necessary to explore a bridge between researchers and integrate decentralized IT resources. Based on this survey, we provide some views concerning IT applying in medical procedures. METHODS: The correlated computer information was collected from a patient with coronary heart disease from early diagnosis, interventional therapy to coronary bypass surgery, until rehabilitation. The medical staffs were investigated from the general status, computer skills, knowledge structure, as well as the acceptance degree of computer application to measure computer skills. RESULTS: The correlated computer information was characterized by large soft variability, a large time span, and various date categories with more dynamic data or three-dimensional data. The investigation of computer knowledge demonstrated that the lack of IT knowledge resulted in poor outcome in their appointed tasks. From the view of self-assessment of IT knowledge, nursing staff and technician exhibited deficiency IT knowledge than doctors, who learned from department training, self-study or internet. Resident physicians and attending physicians were familiar with office system, so they were satisfied with IT skills. In contrary, nurses and majority of high-ranking professor or chief physicians were found difficult to study IT skills. However, most nurses were fond of their office system. The results also demonstrated that people aged 31 -40 years were easy to receive IT system. CONCLUSION: It is very important to enhance the cooperation between computer researchers and medical users for the treatment of coronary atherosclerotic heart disease.

Objective:To analyze the clinical value of different MRI sequences in the diagnosis of elbow trauma in children.Methods:From March 2015 to March 2019, 78 children with elbow joint injury were studied.All patients were examined by X-ray and MRI, including 3D-DESS, TSETIWI, T1-TIRM and FS-PDWI.The differences between the two methods were analyzed.Results:A total of 62 cases of epiphyseal fracture were confirmed by MRI.According to the distribution of Salter Harris fracture: 23 cases of type Ⅱ (37.10%)>17 cases of type Ⅲ (27.42%)>12 cases of type Ⅳ (19.35%)>10 cases of type Ⅰ (16.13%)>0 case of type Ⅴ (0.00%), the difference was statistically significant (χ 2=6.126, P=0.025). Among 78 children of elbow joint injury, 73 cases had bone marrow edema, of which 62 cases were within 1 cm from epiphyseal plate, 11 cases were outside 1 cm from epiphyseal plate, 5 cases were without bone marrow edema, and there was a correlation between bone marrow edema and epiphyseal fracture ( r=0.174, P=0.006). Type Ⅰ-Ⅳ epiphysis injury of elbow joint in children could be accompanied with articular cartilage injury, and 3D-DESS had good effect on articular cartilage injury. Conclusion:Different sequences of MRI have high accuracy in the diagnosis of elbow joint injury in children, which is of great significance in reducing misdiagnosis and missed diagnosis rate of epiphyseal fracture, bone marrow edema and articular cartilage injury in children.

The incidence of coronary heart disease is increasing year by year,with the mortality rate rank-ing first.Early prevention,diagnosis,and treatment are crucial to reduce the mortality rate.Coronary angiography is the gold standard for diagnosing coronary heart disease and is widely used in clinical practice,but it cannot accu-rately reflect the degree of coronary artery disease.Blood flow reserve fraction(FFR)is currently the reference standard for evaluating coronary artery stenosis function.However,it is an invasive examination that is limited in clinical application due to factors such as complex operation,high cost,multiple side effects related to vasodila-tors,and poor reproducibility.In recent years,non-invasive blood flow reserve scores related to imaging have been increasingly applied in clinical practice.Quantitative flow fraction(QFR)and optical coherence tomography based flow reserve fraction(OCT based optical flow ratio OFR)have become research hotspots.OFR,as a relatively new technology,has received increasing attention.Therefore,this article will describe the basic principles,clinical value,and research progress of the above-mentioned technologies.

Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.