Objective： To establish a method of culture and expansion of adult human bone marrow-derived MSCs in vitro and to explore their biological properties. Methods： Mononuclear cells were obtained from 5 mL adult human b...Objective： To establish a method of culture and expansion of adult human bone marrow-derived MSCs in vitro and to explore their biological properties. Methods： Mononuclear cells were obtained from 5 mL adult human bone marrow by density gradient centrifugation with Percoll solution. Adult human MSCs were cultured in Dulbecco＇s Modified Eagle＇s Medium with low glucose （LG-DMEM） containing 10% fetal calf serum at a density of 2×10^5 cell/cm^2. The morphocytology was observed under phase-contrast microscope. The cell growth was measured by MTT method. The flow cytometer was performed to examine the expression of cell surface molecules and cell cycle. The ultrastructure of MSCs was observed under transmission electron microscope. The immunomodulatory functions of MSCs were measured by MTT method. The effects of MSCs on the growth of K562 cells and the dynamic change of HA, IV-C, LN concentration in the culture supernatant of MSCs was also observed. Results： The MSCs harvested in this study were homogenous population and exhibited a spindle-shaped fibroblastic morphology. The cell growth curve showed that MSCs had a strong ability of proliferation. The cells were positive for CD44, while negative for hematopoietic cell surface marker such as CD3, CD4, CD7, CD13, CD14, CD15, CD19, CD22, CD33, CD34, CD45 and HLA-DR, which was closely related to graft versus host disease. Above 90% cells of MSCs were found at G0/G1 phase. The ultrastructure of MSCs indicated that there were plenty of cytoplasmic organelles. Allogeneic peripheral blood lymphocytes proliferation was suppressed by MSCs and the inhibition ratio was 60.68% （P〈0.01）. The suppressive effect was also existed in the culture supernatant of MSCs and the inhibition ratio was 9.00% （P〈0.05）. When lymphocytes were stimulated by PHA, the suppression effects of the culture supernatant were even stronger and the inhibition ratio was 20.91% （P〈0.01）. Compared with the cell growth curve of the K562 ceils alone, the K562 ceils cocultured with MSCs grew slowly and the exponential phase of growth wasn＇t significant. Seeing from the concentration curve, as time passed, the concentration of HA increased quickly, while those of IV-C and LN didn＇t change much. Conclusion： The method for culture and expansion of adult human bone marrow-derived MSCs in vitro has been successfully established in this study. MSCs were a homogenous population that had unique growth phenotype and multilineage potential. Preliminary study proved that it had the abilities of immunomodulatory function, antitumor, hematopoietic supporting and could act as seed cell of tissue engineering.展开更多
Infanticide by males is a common phenomenon in mammals, especially primates, as lactation lasts much longer than gestation in many species. Usually, infanticidal episodes occur soon after group takeovers, and are trad...Infanticide by males is a common phenomenon in mammals, especially primates, as lactation lasts much longer than gestation in many species. Usually, infanticidal episodes occur soon after group takeovers, and are traditionally considered a male reproductive strategy （i.e., support the sexual selection hypothesis, Hrdy, 1974）. To verify the validity of this conception, we observed one group of Franqois＇ langurs in the Nonggang Nature Reserve, China, between August 2003 and July 2004. During the study period, a Francois＇ langur female with the youngest infant in the group was attacked three times by immigrating males, and later disappeared by the third day after the final attack. We suggest that these attacks on the female-infant dyad represent infanticide attempts by males, and may be the cause of the adult female and her infant＇s disappearance. Presumably, that female dispersed with her infant to avoid infanticide and was not killed. Though these observations do not completely verify the sexual selection hypothesis, they are not inconsistent with it.展开更多
The response and recovery mechanisms of forests to damage from freezing rain and snow events are a key topic in forest research and management. However, the relationship between the degree of damage and tree age, i.e....The response and recovery mechanisms of forests to damage from freezing rain and snow events are a key topic in forest research and management. However, the relationship between the degree of damage and tree age, i.e., whether seedlings, young trees, or adult trees are most vulnerable, remains unclear and is rarely reported. We investigated the effect of tree age on the degrees of vegetation damage and subsequent recovery in three subtropical forest types-coniferous, mixed, and broad-leaved —in the Tianjing Mountains, South China, after a series of rare icy rain and freezing snow events in 2008. The results showed that damage and recovery rates were both dependent on tree age, with the proportion of damaged vegetation increasing with age(estimated by diameter at breast height, DBH) in all three forest types and gradually plateauing. Significant variation occurred among forest types. Young trees in the coniferous forest were more vulnerable than those in the broad-leaved forest. The type of damage also varied with tree age in different ways in the three forest types. The proportion of young seedlings that were uprooted(the most severe type of damage) was highest in the coniferous forest. In the mixed forest, young trees were significantly more likely to be uprooted than seedlings and adult trees, while in the broad-leaved forest, the proportion of uprooted adult trees was significantly higher than that of seedlings and young trees. There were also differences among forest types in how tree age affected damage recovery. In the coniferous forest, the recovery rate of trees with broken trunks or crowns(DBH > 2.5 cm) increased with tree age. However, in the mixed and broad-leaved forests, no obvious correlation between the recovery rate of trees with broken trunks or crowns and tree age was observed. Trees with severe root damage did not recover; they were uprooted and died. In these forests, vegetation damage and recovery showed tree age dependencies, which varied with tree shape, forest type, and damage type. Understanding this dependency will guide restoration after freezing rain and snow disturbances.展开更多
基金Supported by the grant from Lanzhou Command Medical Research Foundation (No. LXH-2005013).
文摘Objective： To establish a method of culture and expansion of adult human bone marrow-derived MSCs in vitro and to explore their biological properties. Methods： Mononuclear cells were obtained from 5 mL adult human bone marrow by density gradient centrifugation with Percoll solution. Adult human MSCs were cultured in Dulbecco＇s Modified Eagle＇s Medium with low glucose （LG-DMEM） containing 10% fetal calf serum at a density of 2×10^5 cell/cm^2. The morphocytology was observed under phase-contrast microscope. The cell growth was measured by MTT method. The flow cytometer was performed to examine the expression of cell surface molecules and cell cycle. The ultrastructure of MSCs was observed under transmission electron microscope. The immunomodulatory functions of MSCs were measured by MTT method. The effects of MSCs on the growth of K562 cells and the dynamic change of HA, IV-C, LN concentration in the culture supernatant of MSCs was also observed. Results： The MSCs harvested in this study were homogenous population and exhibited a spindle-shaped fibroblastic morphology. The cell growth curve showed that MSCs had a strong ability of proliferation. The cells were positive for CD44, while negative for hematopoietic cell surface marker such as CD3, CD4, CD7, CD13, CD14, CD15, CD19, CD22, CD33, CD34, CD45 and HLA-DR, which was closely related to graft versus host disease. Above 90% cells of MSCs were found at G0/G1 phase. The ultrastructure of MSCs indicated that there were plenty of cytoplasmic organelles. Allogeneic peripheral blood lymphocytes proliferation was suppressed by MSCs and the inhibition ratio was 60.68% （P〈0.01）. The suppressive effect was also existed in the culture supernatant of MSCs and the inhibition ratio was 9.00% （P〈0.05）. When lymphocytes were stimulated by PHA, the suppression effects of the culture supernatant were even stronger and the inhibition ratio was 20.91% （P〈0.01）. Compared with the cell growth curve of the K562 ceils alone, the K562 ceils cocultured with MSCs grew slowly and the exponential phase of growth wasn＇t significant. Seeing from the concentration curve, as time passed, the concentration of HA increased quickly, while those of IV-C and LN didn＇t change much. Conclusion： The method for culture and expansion of adult human bone marrow-derived MSCs in vitro has been successfully established in this study. MSCs were a homogenous population that had unique growth phenotype and multilineage potential. Preliminary study proved that it had the abilities of immunomodulatory function, antitumor, hematopoietic supporting and could act as seed cell of tissue engineering.
基金supported by National Nature Science Foundation of China (31172122)Guangxi Nature Science Foundation (2012GXNSFAA053039)
文摘Infanticide by males is a common phenomenon in mammals, especially primates, as lactation lasts much longer than gestation in many species. Usually, infanticidal episodes occur soon after group takeovers, and are traditionally considered a male reproductive strategy （i.e., support the sexual selection hypothesis, Hrdy, 1974）. To verify the validity of this conception, we observed one group of Franqois＇ langurs in the Nonggang Nature Reserve, China, between August 2003 and July 2004. During the study period, a Francois＇ langur female with the youngest infant in the group was attacked three times by immigrating males, and later disappeared by the third day after the final attack. We suggest that these attacks on the female-infant dyad represent infanticide attempts by males, and may be the cause of the adult female and her infant＇s disappearance. Presumably, that female dispersed with her infant to avoid infanticide and was not killed. Though these observations do not completely verify the sexual selection hypothesis, they are not inconsistent with it.
基金financially supported by the National Natural Science Foundation of China(31300401,31030015,31100402)the Forestry Science and Technology Innovative Foundation of Guangdong Province(2008KJCX012,2009KJCX015)+1 种基金the Guangdong Natural Science Foundation(S2012040007896)the Fundamental Research Funds for the Central Universities
文摘The response and recovery mechanisms of forests to damage from freezing rain and snow events are a key topic in forest research and management. However, the relationship between the degree of damage and tree age, i.e., whether seedlings, young trees, or adult trees are most vulnerable, remains unclear and is rarely reported. We investigated the effect of tree age on the degrees of vegetation damage and subsequent recovery in three subtropical forest types-coniferous, mixed, and broad-leaved —in the Tianjing Mountains, South China, after a series of rare icy rain and freezing snow events in 2008. The results showed that damage and recovery rates were both dependent on tree age, with the proportion of damaged vegetation increasing with age(estimated by diameter at breast height, DBH) in all three forest types and gradually plateauing. Significant variation occurred among forest types. Young trees in the coniferous forest were more vulnerable than those in the broad-leaved forest. The type of damage also varied with tree age in different ways in the three forest types. The proportion of young seedlings that were uprooted(the most severe type of damage) was highest in the coniferous forest. In the mixed forest, young trees were significantly more likely to be uprooted than seedlings and adult trees, while in the broad-leaved forest, the proportion of uprooted adult trees was significantly higher than that of seedlings and young trees. There were also differences among forest types in how tree age affected damage recovery. In the coniferous forest, the recovery rate of trees with broken trunks or crowns(DBH > 2.5 cm) increased with tree age. However, in the mixed and broad-leaved forests, no obvious correlation between the recovery rate of trees with broken trunks or crowns and tree age was observed. Trees with severe root damage did not recover; they were uprooted and died. In these forests, vegetation damage and recovery showed tree age dependencies, which varied with tree shape, forest type, and damage type. Understanding this dependency will guide restoration after freezing rain and snow disturbances.
