1.Establishment of the EMT model of CoCl2 -induced NRK-52E cells
Lei Ni ; Qingqing Sun ; Jiangrui Cheng ; Wei Wei ; Chun Wang
Acta Universitatis Medicinalis Anhui 2025;60(10):1879-1886
Objective:
To establish an in vitro cell model of Cobalt dichloride(CoCl2)-induced epithelial-mesenchymal transition(EMT) in rat renal tubular epithelial cells(NRK-52E).
Methods:
NRK-52E cells were cultured in vitro and randomly divided into a blank control group(NC group) and a CoCl2 treatment group. The CoCl2 treatment group underwent 1, 2, 3, and 4 cycles of treatment, with each cycle consisting of CoCl2 treatment for 9 h followed by recovery in CoCl_2-free medium for 3 h. The optimal concentration and time of CoCl_2-induced EMT were screened using the CCK-8 assay. Morphological changes in cells were observed using light microscopy and phalloidin staining. The expression levels of EMT marker proteins were detected by Western blot and immunofluorescence.
Results:
Compared with the NC group , stimulation by 100 μmol/L CoCl2 for 48 h significantly induced apoptosis (P < 0. 01) , meeting the requirements for subsequent experiments. Western blot results showed that the expression of hypoxia-inducible factor-1 α (HIF-1α ) and α-smooth muscle actin ( α-SMA) significantly increased in the 3-cycle group treated with 100 μmol/L CoCl2 for 9 h followed by recovery for 3 h ( P < 0. 001) , indicating the most pronounced fibrotic response. Observations under light microscopy and rhodamine-labeled phalloidin staining revealed that the morphological changes and cytoskeletal rearrangement of NRK-52E cells were most significant in the 3-cycle group treated with 100 μmol/L CoCl2 for 9 h followed by recovery for 3 h , demonstrating the best model stability. Immunofluorescence results showed that compared with the NC group , the fluorescence intensity of the fibrous matrix protein Collagen I significantly increased in the 3-cycle group treated with 100 μmol/L CoCl2 for 9 h followed by recovery for 3 h ( P < 0. 001) .
Conclusion
The protocol involves treating NRK-52E cells with 100 μmol/L CoCl2 for 9 h , following 3 h of recovery in CoCl2 -free medium. Repeating this cycle three times can establish an in vitro EMT model.
2.GC-MS Analysis on Essential Oil and Liposoluble Constituents of Lepidium Meyenii Walp.
Lihong TAN ; Hailin RAN ; Jiangrui WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(7):1234-1238
In order to provide scientific basis for quality evaluation of Lepidium meyenii Walp.in Sichuan province,the extracted constituents of essential oil and petroleum ether were analyzed by gas chromatography-mass spectrometry (GC-MS),respectively.The volatile oil and fat soluble components were extracted by the steam distillation and petroleum ether ultrasonic.They were isolated and identified by GC-MS.The structures were identified in combination with database search system of MS.The relative content of every compound was determined by normalization method.The results showed that a total of 22 compounds were identified in volatile oil (98.85%) and 40 compounds were identified in liposoluble constituents (73.99%).There were significant differences of components and relative percentage contents of volatile oil and fat-soluble components.Dodecane,quinoline,3-methoxybenzaldehyde,palmitic acid methyl ester and ethyl linoleate were the same compositions.Comparative analysis was conducted on volatile oil and liposoluble constituents by GC-MS for the first time.The results provided references for herbal resource investigation and quality standard evaluation of Maca.
3.Investigation and analysis of vigilance and attention of the ship crew at different operational posts
Cun WEI ; Yunxia WANG ; Jiangrui ZHOU ; Guanghui DENG ; Xiaoli CHEN ; Xinghua SHEN
Chinese journal of nautical medicine and hyperbaric medicine 2015;22(4):286-289
Objective To investigate effects of different working environment on the vigilance and attention of the ship crew.Methods The times of correct response of the ship crew at different posts to simultaneous sound-light reaction,progress in the sound-light reaction time,the time on targets and the time out of track of targets were detected by using attention distribution measuring instrument (EP707A) and attention focus measuring instrument (EP711).Results There were significant differences in attention focusing for the ship crew at different posts (F =3.78,P =0.006).Different working environment had significant effects on attention focusing of the ship crew with more than 3 years of service at sea.Conclusions The ship crew working in the electromechanical and ammunition departments had relatively poor attention focusing.Three to five years of service onboard the ship was an important episode,when attention focusing was affected or most likely to be affected.
4.Investigation and analysis of vigilance and attention of the ship crew at different operational posts
Cun WEI ; Yunxia WANG ; Jiangrui ZHOU ; Guanghui DENG ; Xiaoli CHEN ; Xinghua SHEN
Chinese journal of nautical medicine and hyperbaric medicine 2015;22(4):286-289
Objective To investigate effects of different working environment on the vigilance and attention of the ship crew.Methods The times of correct response of the ship crew at different posts to simultaneous sound-light reaction,progress in the sound-light reaction time,the time on targets and the time out of track of targets were detected by using attention distribution measuring instrument (EP707A) and attention focus measuring instrument (EP711).Results There were significant differences in attention focusing for the ship crew at different posts (F =3.78,P =0.006).Different working environment had significant effects on attention focusing of the ship crew with more than 3 years of service at sea.Conclusions The ship crew working in the electromechanical and ammunition departments had relatively poor attention focusing.Three to five years of service onboard the ship was an important episode,when attention focusing was affected or most likely to be affected.
5.Clinical study on sequential treatment of newly diagnosed de novo acute myeloid leukemia patients with IA and low-dose HA combined with G-CSF regimens as remission induction therapy
Zhengjin ZHENG ; Shaoyuan WANG ; Langhui ZHANG ; Qingxiao SONG ; Jiangrui GUO
Journal of Leukemia & Lymphoma 2012;21(8):481-483
Objective To evaluate the efficacy and safety of sequential treatment of newly diagnosed de novo acute myeloid leukemia (AML) patients with IA and low-dose HA combined with G-CSF regimens as remission induction therapy.Methods Fifty-seven patients with AML were enrolled,which marrow biopsy was hypocellular or active proliferation on the third day from the end of the first course with IA regimen.32 cases of them received the second course with low-dose HA combined with G-CSF regimen,compared with other 25 cases received the second course with another IA regimen.Clinical manifestations,blood count,blood biochemical parameters and bone marrow smears were measured during the courses.Results In study group,21 of 32 cases reached CR,4 PR,and 11 of 20 cases reached CR,2 PR in control group.Overall remission rate (ORR) was higher in study group than that in control group (78.1% vs 52.0 %,P =0.038).Both median duration of agranulocytosis and median time for PLT to reach 50×109/L from the lowest were shorter in study group than those in control group (9.5 d vs 28.0 d,U=32.5,P< 0.001; 11 d vs 19 d,U=193.0,P=0.001).Component transfusion,not only RBC but PLT,decreased in study group,compared with control group (8 U vs 16 U,U =206.5,P =0.002; 20 U vs 60 U,U =149,P < 0.001).Median durable time of antibiotic intravenous injection was shorter in study group than that in control group (14 d vs 21 d,U=249.5,P=0.015).Visceral hemorrhage rate reduced in study group,compared with control group (x2 =3.90,P =0.048).Conclusion IA and low-dose HA combined with G-CSF regimens sequential treatment as remission induction therapy for newly diagnosed de novo AML patients is effective and well tolerated.


Result Analysis
Print
Save
E-mail