Objective To establish an aging model of mesenchymal stem cells (MSCs) and to investigate aging related biological mechanism for the purpose of studying the senesence of MSCs .Methods MSCs were separated and purified from human placenta, and the cells of the third passage(P3-MSCs) were cultured in the medium for 2 hours, then 100,200 and 300 μmol/L hydrogen peroxide ( H2 O2 ) was added to the cells for 2 hours to establish the MSCs aging model in vitro. Biological characteristics of aging MSCs were evaluated by cell cycle assay and senescence associated β-galactosidase staining.The expression of p16,p21 and p53 genes was further measured using quantitative real-time PCR (RT-PCR).Re-sults Compared with the control , the number of MSCs treated with 200μmol/L H2 O2 for 2 hours was significantly decreased and the cells displayed less adipogenic ,osteogenic and chondrogenic differentiation .Moreover ,after exposure to 200 μmol/L H2 O2 , the majority of the cells were in the G 0/G1 phase as showed by cell cycle analysis .The percentage of senescence-associated β-galactosidase-positive cells was increased , and the expression of p 16 , p21 and p53 mRNA and protein was significantly increased.Conclusion The results of this study has demonstrated that the H 2 O2 (200 μmol/L) can be used to establish the aging model of MSCs in vitro, and the cellular phenotypic alteration may attribute to the cell cycle associated gene expression (p16, p21, and p53).

BACKGROUND:Placenta is a valuable source of mesenchymal stem cels for stem cel therapy and future application in the field of regenerative medicine. However, conventional methods cannot acquire a large amount of purified human placenta-derived mesenchymal stem cels. Here, we present a new method for isolating human placenta-derived mesenchymal stem cels suitable for banking strategies and for future clinical applications. OBJECTIVE:To analyze the biological characteristics of human placenta-derived mesenchymal stem cels cultured by tissue dissociating and colagenase digestion. METHODS: Human placenta-derived mesenchymal stem cels were obtained from human placenta by tissue dissociating and colagenase digestion method. Immunophenotype was analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated byin vitro adipogenic, osteogenic and chondrogenic induction as wel. RESULTS AND CONCLUSION:Human placenta-derived mesenchymal stem cels could be passaged stablyin vitro. Furthermore, the cels expressed CD73, CD90, CD105, but were negative for the markers of CD11b, CD19, CD34, CD45, and HLA-DR. Human placenta-derived mesenchymal stem cels proliferated actively and began to grow logarithmicaly at days 3-5 folowed by a plateau period at day 6. In addition, the isolated cels could be induced into adipocytes, osteocytes, chondrocytesin vitro. In a word, the results of this study demonstrated that the tissue dissociating and colagenase digestion method is an efficient method for obtaining a large amount of human placenta-derived mesenchymal stem cels that can be stably cultured in vitro and have strong proliferative ability.

Objective To investigate the effect of cartilage acetabular ,glenoid labrum and acetabular tissue on closed reduction of developmental dysplasia of hip ( DDH ) by analyzing the outcomes of the ultrasound measurement on DDH before and after Pavik Harness treatment . Methods Graf method was used in ultrasound measurement before the treatment . Position of cartilaginous acetabulum and glenoid labrum and displacement direction of femoral head in 31 patients with hip dislocations ( type D ,type Ⅲ ,typeⅣ ,36 hips) were recorded . T he follow‐up testing with ultrasound measurement on the inside of the hip were performed after 1 －2 weeks of Pavik Harness treatment . M ediolateral dimension ,anteroposterior dimension and the area of maximum section of acetabular tissue were measured . Reduction effects were evaluated . Results T he success rate of reduction of dislocation hips with cartilaginous acetabulum on the head orientation was obviously higher than that on the foot orientation( P ＜0 .000 1) . T he success rate of reduction of dislocation hips with glenoid labrum on the head orientation was obviously higher than that on the foot orientation ( P ＝ 0 .001 6 ) . T here was a statistically significant difference in the success rate of different types of hip dislocation ( type D , type Ⅲ , type Ⅳ ) ( P＝ 0 .002 7 ) . T here were statistically significant differences in mediolateral dimension ,anteroposterior dimension and the area of maximum section of acetabular tissue between the reduction group and the group without dislocation ,the irreducible group and the group without dislocation ,the irreducible group and the reduction group ( P ＜ 0 .01) . Conclusions Ultrasonography is of great significance in the diagnosis and treatment of children with DDH . T he position of hip cartilaginous acetabulum and glenoid labrum ,types of hip dislocation and the volume of acetabular tissue have important effects on the success of the closed reduction .

Objective:To analyze the contagiousness and secondary attack rate of 2019 novel coronavirus in cluster epidemics in Guangzhou and provide evidence for the prevention and control of COVID-19.Methods:All the individuals identified to be infected with 2019-nCoV in Guangzhou, including confirmed cases and asymptomatic cases, were included and classified as imported cases and local cases. The first case of each cluster epidemic was defined as index case, and the number of subsequent infections was calculated to evaluate the contagiousness and secondary attack rate of 2019 novel coronavirus in the shortest incubation period of 1-3 days.Results:As of 18 February, 2020, a total of 349 cases of 2019-nCoV infection, including 339 confirmed cases (97.13 %) and 10 asymptomatic cases (2.87 %) were reported in Guangzhou. There were 68 clusters involving 217 2019-nCoV infection cases (210 confirmed cases and 7 asymptomatic cases). The median number of subsequent infections caused by an index case in a cluster epidemic was 3, among which 2 were confirmed cases and 1 was asymptomatic cases, respectively. The average number of contagiousness was 2.18 in shorted incubation period of 1-3 days (The average number of infected cases were 2.18 cases by the index case in a cluster epidemic), the average infection number in family members was 1.86, and the infection ratio of family member transmission was 85.32 % (1.86/2.18). The secondary attack rate in close contacts with shortest incubation period of 1-3 days was 17.12 %-18.99 %, the secondary attack rate in family members was 46.11 %-49.56 %. Conclusions:The cluster epidemic of COVID-19 in Guangzhou mainly occurred in families, the contagiousness was high. It is necessary to strengthen the prevention and control to reduce the community transmission of COVID-19.