1.TAB182 promotes DNA homologous recombination repair by stabilizing RPA2 mRNA
Gang LI ; Shuting LAI ; Yang HAN ; Chenjun BAI ; Hua GUAN ; Shanshan GAO ; Pingkun ZHOU
Chinese Journal of Radiological Medicine and Protection 2022;42(4):241-247
Objective:To investigate the regulating molecules and acting mechanism of TAB182 in HR pathway.Methods:TAB182 in human breast cancer MCF-7 cells was knocked down by shRNA strategy, the TAB182 knockdown MCF-7 as the TAB182 knockdown group, and the MCF-7 cell using the shRNA negative control as the TAB182 negative control group. RNA sequencing and qRT-PCR were performed to screen and verify the differentially expressed genes of HR pathway related to TAB182 depression. Western blot was used to detect protein expression. Immunofluorescence staining of nuclear RAD51 and BrdU was used to check the 3′ ssDNA formation by the end resection. The cell cycle arrest and apoptosis were measured by flow cytometry. Cloning formation assay was used to evaluate the sensitivity TAB182-knockdown cells to radiation.Results:Both quantitative RNA sequencing and qRT-PCR assays showed that TAB182-knockdown significantly decreased the mRNA expression of RPA2( t=17.97, P<0.05). Compared with the TAB182 negative control group, the protein level of RPA2, the number of RAD51 foci, and the 3′ ssDNA-binding nuclear protein marker BrdU in TAB182-knockdown cells were significantly reduced. At 4, 8, and 12 h after actinomycin D treatment, the attenuation of RPA2 mRNA in the TAB182-knockdown cells was accelerated ( t=5.37, 3.79, 3.69, P<0.05). Compared with the TAB182 negative control group, the radiosensitivity and radiation-induced apoptosis in the TAB182-knockdown group were increased ( t=3.48, 11.05, P<0.05), and at 24 h after irradiation, the cell cycle block time was prolonged ( t=8.40, P<0.01). Conclusions:TAB182 plays a role in maintaining RPA2 mRNA stability, thereby promoting HR repair. TAB182 knockdown cells are highly sensitive to ionizing radiation.
2.Influencing factors of the TEG maximum amplitude reduction in emergency trauma patients
Changfu JI ; Dong LAI ; Yan TIAN ; Fei LAI ; Shuting LIN ; Min LONG ; Fangyuan LI ; Fangling LIAN
Chinese Journal of Blood Transfusion 2024;37(6):638-642
【Objective】 To investigate the causes of abnormal decrease in maximum amplitude(MA) of thromboelastography(TEG) and its effect on prognosis by monitoring the changes of coagulation-related indexes in emergency trauma patients. 【Methods】 A total of 319 cases of trauma patients admitted to our hospital from September 2020 to September 2023 were retrospectively analyzed, and the coagulation-related indexes of 0 h and 24 h after admission were observed. According to the MA results, they were divided into normal MA group(>50 mm) and reduced MA group(≤50 mm) to compare the hemoglobin(Hb), platelets count(Plt), activated partial thromboplastin time(APTT), prothrombin time(PT), fibrinogen(Fib), thrombin time(TT), D-dimer(D-D), coagulation reaction time(R), clot formation kinetics(Angle), 30 min clot dissolution rate(Ly30), MA, thrombine-antithrombin complex(TAT) and plasminase-α2 plasminase inhibitor complex(PIC). The correlation between MA and fibrinolysis indexes in 319 trauma patients was analyzed. According to whether tranexamic acid(TXA) was used, the reduced MA group was divided into a TXA group and a non-drug group. The differences in the change of the above coagulation-related indexes, mortality rate and changes in blood product dosage were compared between the two groups. 【Results】 Compared with the normal MA group, Hb, Plt, Fib, diastolic blood pressure and GCS scores decreased, while heart rate, ISS score and mortality increased significantly in the reduced MA group(P<0.05). The R, PT and TT were prolonged significantly(P<0.05), and PIC and D-D increased significantly(P<0.05) in the reduced MA group. Correlation analysis found that MA had no correlation with Ly30, TAT and APTT, but was correlated with Angle(r=0.803), Plt(r=0.544), Fib(r=0.581), PIC(r=-0.443) and D-D(r=-0.343). Compared with the non-drug group, the change of Angle, MA and FIB in the TXA group increased significantly(P<0.05), while the change of PIC decreased(P<0.05). Cryoprecipitate and platelet transfusion in the TXA group reduced significantly(P<0.05), and red blood cell transfusion had a decreasing trend, but the difference was not significant(P>0.05). The mortality rate in the TXA group was reduced significantly(P<0.05). 【Conclusion】 Hyperfibrinolysis may be an important factor in the abnormal decrease of MA in emergency trauma patients. Treatment with TXA can improve its effect on MA, and reduce the transfusion of blood products and the patient mortality.
3.Transcriptome analysis reveals the role of withering treatment in flavor formation of oolong tea (Camellia sinensis).
Chen ZHU ; Shuting ZHANG ; Chengzhe ZHOU ; Biying SHI ; Linjie HUANG ; Yuling LIN ; Zhongxiong LAI ; Yuqiong GUO
Chinese Journal of Biotechnology 2022;38(1):303-327
Oolong tea is a semi-fermented tea with strong flavor, which is widely favored by consumers because of its floral and fruity aroma as well as fresh and mellow taste. During the processing of oolong tea, withering is the first indispensable process for improving flavor formation. However, the molecular mechanism that affects the flavor formation of oolong tea during withering remains unclear. Transcriptome sequencing was used to analyze the difference among the fresh leaves, indoor-withered leaves and solar-withered leaves of oolong tea. A total of 10 793 differentially expressed genes were identified from the three samples. KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in flavonoid synthesis, terpenoid synthesis, plant hormone signal transduction and spliceosome pathways. Subsequently, twelve differentially expressed genes and four differential splicing genes were identified from the four enrichment pathways for fluorescence quantitative PCR analysis. The results showed that the expression patterns of the selected genes during withering were consistent with the results in the transcriptome datasets. Further analysis revealed that the transcriptional inhibition of flavonoid biosynthesis-related genes, the transcriptional enhancement of terpenoid biosynthesis-related genes, as well as the jasmonic acid signal transduction and the alternative splicing mechanism jointly contributed to the flavor formation of high floral and fruity aroma and low bitterness in solar-withered leaves. The results may facilitate better understanding the molecular mechanisms of solar-withering treatment in flavor formation of oolong tea.
Camellia sinensis/genetics*
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Gene Expression Profiling
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Plant Leaves
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Plant Proteins/metabolism*
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Taste
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Tea
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Transcriptome/genetics*