Establishment and Evaluation of Intestinal Injury Model of Mouse Acute Graft Versus Host Disease Based on An Organoid Technology / 中国实验血液学杂志

OBJECTIVE@#To establish an intestinal organoid model that mimic acute graft versus host disease (aGVHD) caused intestinal injuries by using aGVHD murine model serum and organoid culture system, and explore the changes of aGVHD intestine in vitro by advantage of organoid technology.@*METHODS@#20-22 g female C57BL/6 mice and 20-22 g female BALB/c mice were used as donors and recipients for bone marrow transplantation, respectively. Within 4-6 h after receiving a lethal dose (8.0 Gy) of γ ray total body irradiation, a total of 0.25 ml of murine derived bone marrow cells (1×107/mice, n=20) and spleen nucleated cells (5×106/mice, n=20) was infused to establish a mouse model of aGVHD (n=20). The aGVHD mice were anesthetized at the 7th day after transplantation, and the veinal blood was harvested by removing the eyeballs, and the serum was collected by centrifugation. The small intestinal crypts of healthy C57BL/6 mice were harvested and cultivated in 3D culture system that maintaining the growth and proliferation of intestinal stem cells in vitro. In our experiment, 5%, 10%, 20% proportions of aGVHD serum were respectively added into the organoid culture system for 3 days. The formation of small intestinal organoids were observed under an inverted microscope and the morphological characteristics of intestinal organoids in each groups were analyzed. For further evaluation, the aGVHD intestinal organoids were harvested and their pathological changes were observed. Combined with HE staining, intestinal organ morphology evaluation was performed. Combined with Alcian Blue staining, the secretion function of aGVHD intestinal organoids was observed. The distribution and changes of Lgr5+ and Clu+ intestinal stem cells in intestinal organoids were analyzed under the conditions of 5%, 10% and 20% serum concentrations by immunohistochemical stainings.@*RESULTS@#The results of HE staining showed that the integrity of intestinal organoids in the 5% concentration serum group was better than that in the 10% and 20% groups. The 5% concentration serum group showed the highest number of organoids, the highest germination rate and the lowest pathological score among experimental groups, while the 20% group exhibited severe morphological destruction and almost no germination was observed, and the pathological score was the highest among all groups(t=3.668, 4.334,5.309,P<0.05). The results of Alican blue staining showed that the secretion function of intestinal organoids in serum culture of aGVHD in the 20% group was weaker than that of the 5% group and 10% of the organoids, and there was almost no goblet cells, and mucus was stainned in the 20% aGVHD serum group. The immunohistochemical results showed that the number of Lgr5+ cells of intestinal organoids in the 5% group was more than that of the intestinal organoids in the 10% aGVHD serum group and 20% aGVHD serum group. Almost no Clu+ cells were observed in the 5% group. The Lgr5+ cells in the 20% group were seriously injuried and can not be observed. The proportion of Clu+ cells in the 20% group significantly increased.@*CONCLUSION@#The concentration of aGVHD serum in the culture system can affect the number and secretion function of intestinal organoids as well as the number of intestinal stem cells in organoids. The higher the serum concentration, the greater the risk of organoid injury, which reveal the characteristics of the formation and functional change of aGVHD intestinal organoids, and provide a novel tool for the study of intestinal injury in aGVHD.

Secondary metabolites from the dandelion-derived endophytic fungus Epicoccum sorghinum 1-2 / 药学学报

The secondary metabolites from the dandelion-derived Epicoccum sorghinum 1-2 were isolated by silica gel and Sephadex gel column chromatography, and semi-preparative high performance liquid chromatography (HPLC). Their structures were identified by comprehensive NMR and MS methods. Their antibacterial activities were determined by filter paper method. Finally, seven compounds were isolated and identified from the fermentation product of E. sorghinum 1-2, including (4R*,5R*,6S*)-4,5-dihydroxy-6-(6'-methylsalicyloxy)-2-methoxymethyl-2-cyclohexen-l-one (1), (4R*,5R*,6S*)-4,5-dihydroxy-6-(6′-methylsalicyloxy)-2-methyl-2-cyclohexen-1-one (2), (4R,5R,6S)-4,5-dihydroxy-6-(6'-methylsalicyloxy)-2-hydroxymethyl-2-cyclohexen-1-one (3), (-)-gabosine E (4), theobroxide (5), 3-chlorogentisyl alcohol (6), and 3-hydroxybenzyl alcohol (7), of which 1-5 are epoxydons, and 6 and 7 are phenolics. Compounds 1 and 2 are new structures reported for the first time. Compound 6 showed significant antibacterial activity against Staphylococcus aureus.

The burden of hip fracture and related factors in Chinese middle-aged and elderly population / 中华骨科杂志

Objective:To estimate the burden of hip fractures and related factors among the Chinese middle-aged and elderly population.Methods:Based on the data of China Health and Retirement Longitudinal Study (CHARLS) in 2015, the burden of hip fractures among Chinese middle-aged and elderly population was evaluated as the years lived with disability (YLD) rate. The relationship between gender, age (45-49, 50-59, 60-69 and ≥70 years old), educational level (no education, elementary school, junior high school, high school and above), region (urban, rural), gross domestic production (GDP) per capita (low, medium and high) and geographic area (northern, eastern, south-central, northwest and southwest regions) and the YLD rate of hip fractures were analyzed, respectively.Results:Excluding items with missing basic information and hip fracture data, a total of 17,830 subjects (8,405 males and 9,425 females) were included in the study with average age 61.6±9.8 years (range 45 to 105 years). The average age in male was 61.9±9.6 years, and that in female was 61.2±10.1 years. The prevalence of hip fractures was 2.3% (410/17,830). The total YLD rate was [694/100,000 (95% Uncertainty Interval ( UI): 462/100,000, 989/100,000)]. The YLD rates were similar between male [693/100,000 (95% UI: 461/100,000, 988/100,000)] and female [696/100,000 (95% UI: 463/100,000, 992/100,000)] subgroups. The YLD rate of hip fractures was increasing with age, which reached at maximum of 1,155/100,000 (95% UI: 769/100,000, 1,646/100,000) for participants aged 70 years or above. The rate was gradually decreased with the upgrade of the educational level. The participants with high school education and above reached the lowest of 434/100,000 (95% UI: 289/100,000, 619/100,000). In addition, the YLD rate of hip fractures in rural areas [721/100,000 (95% UI: 480/100,000, 1,027/100,000)] was higher than that in urban areas [650/100,000 (95% UI: 433/100,000, 926/100,000)]. The YLD rate in areas with higher GDP per capita [545/100,000 (95% UI: 363/100,000, 777/100,000)] was lower than that in areas with lower GDP per capita [761/100,000 (95% UI: 506/100,000, 1,084/100,000)]. Moreover, the participants living in the Northwest region were with the highest YLD rate of hip fractures [1,056/100,000 (95% UI: 703/100,000, 1,506/100,000)], followed by the Southwest region [887/100,000 (95% UI: 590/ 100,000, 1,264/100,000)] and the Northeast region [317/100,000 (95% UI: 211/100,000, 452/100,000)]. Conclusion:Hip fractures exerted heavy burdens on the Chinese middle-aged and elderly population. The YLD rate of hip fractures varied according to geographical regions, greater age, rural areas, low educational levels and low GDP per capita. These related factors could affect the disease burden of hip fractures in China.