Role of ASXLI loss in biological functions of HEL cells / 中国药理学通报

Aim To reveal the underlying mechanisms of the co-occurrence of ASXLI and JAK2ymr mutation by using human leukemia cell line HEL that carried homozygous /4A2V617F mutation in the elucidation of the role of ASXLI loss of function mutation in myeloproliferative neoplasms, so as to provide an important model for investigating the role of ASXLI mutation in myeloproliferative neoplasms at the cellular level. Methods HEL cell line with ASXLI knockout ( HEL-AKO) was established by using CRISPR/Cas9-mediated gene editing. And a series experiments based were preformed to verify the effect of ASXLI on HEL cell proliferation, clone formation and chemosensitivity. Results HEL-AKO cell line was successfully established, confirmed by sequencing results. We found that the loss of ASXLI could inhibit proliferation and induce cell cycle arrest at the G2/M phase. The colony-form- ing capacity of HEL-AKO cells was also markedly inhibited. Moreover, the HEL-AKO had higher cloning efficiency than HEL Control after ruxolitinib treatment. Conclusions Loss of function of ASXLI has an impact on cell biological function of HEL. Therefore, HEL- AKO cell line can be used to further explore the biological contribution of concomitant ASXLI in /4A2V617F mutated MPN.

Rapid discovery and identification of the anti-inflammatory constituents in Zhi-Shi-Zhi-Zi-Chi-Tang / 中国天然药物

The anti-inflammatory active ingredients of Zhi-Shi-Zhi-Zi-Chi-Tang (ZSZZCT), a traditional Chinese medicine formula, were predicted and identified using an approach based on activity index, LC-MS, semi-preparative LC and NMR. Firstly, the whole extract of ZSZZCT was analyzed using liquid chromatography-quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) and liquid chromatography - ion trap mass spectrometry (LC-IT-MS), 79 constituents were detected and 39 constituents were identified unambiguously or tentatively. Subsequently, the whole extract of the formula was separated into multiple components and the activity index method was used to calculate index values of the 79 constituents by integrating the chemical and pharmacological information of multiple components. Four polymethoxyl flavones were predicted as the major active constituents according to the activity index values. Furthermore, three polymethoxyl flavones were prepared using the strategy with semi-preparative LC guided by LC-MS, and their anti-inflammatory activities were validated. The results show that three polymethoxyl flavones with higher positive index values, i.e., 3, 5, 6, 7, 8, 3', 4'-heptamethoxyflavone, 3-hydroxynobiletein and tangeretin had significant anti-inflammatory effects. In conclusion, the predicted results indicated that the activity index method is feasible for the accurate prediction of active constituents in TCM formulae.

The body weight loss during acute exposure to high-altitude hypoxia in sea level residents / 生理学报

Weight loss is frequently observed after acute exposure to high altitude. However, the magnitude and rate of weight loss during acute exposure to high altitude has not been clarified in a controlled prospective study. The present study was performed to evaluate weight loss at high altitude. A group of 120 male subjects [aged (32±6) years] who worked on the construction of the Golmud-Lhasa Railway at Kunlun Mountain (altitude of 4 678 m) served as volunteer subjects for this study. Eighty-five workers normally resided at sea level (sea level group) and 35 normally resided at an altitude of 2 200 m (moderate altitude group). Body weight, body mass index (BMI), and waist circumference were measured in all subjects after a 7-day stay at Golmud (altitude of 2 800 m, baseline measurements). Measurements were repeated after 33-day working on Kunlun Mountain. In order to examine the daily rate of weight loss at high altitude, body weight was measured in 20 subjects from the sea level group (sea level subset group) each morning before breakfast for 33 d at Kunlun Mountain. According to guidelines established by the Lake Louise acute mountain sickness (AMS) consensus report, each subject completed an AMS self-report questionnaire two days after arriving at Kunlun Mountain. After 33-day stay at an altitude of 4 678 m, the average weight loss for the sea level group was 10.4% (range 6.5% to 29%), while the average for the moderate altitude group was 2.2% (-2% to 9.1%). The degree of weight loss (Δ weight loss) after a 33-day stay at an altitude of 4 678 m was significantly correlated with baseline body weight in the sea level group (r=0.677, P<0.01), while the correlation was absent in the moderate altitude group (r=0.296, P>0.05). In the sea level subset group, a significant weight loss was observed within 20 d, but the weight remained stable thereafter. AMS-score at high altitude was significantly higher in the sea level group (4.69±2.48) than that in the moderate altitude group (2.97±1.38), and was significantly correlated with baseline body weight. These results indicate that (1) the person with higher body weight during stay at high altitude loses more weight, and this is more pronounced in sea level natives when compared with that in moderate altitude natives; (2) heavier individuals are more likely to develop AMS than leaner individuals during exposure to high-altitude hypoxia.