3.Characteristics of bax, bcl-2 and p53 gene expression in children and fetal skin at different developmental stages
Wei CHEN ; Xiaobing FU ; Shili GE ; Gang ZHOU ; Duyin JIANG ; Tongzhu SUN ; Bing HAN ; Zhiyong SHENG
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To investigate gene expression of bax, bcl-2 and p53 in fetal skin at different gestational ages and children skin in order to explore their potentially biological significance. METHODS: Apoptosis in skin specimens was determined by terminal deoxynucleotidy transferase mediated dUTP-biotin nick-end labeling technique (TUNEL). Gene expressions of bax, bcl-2 and p53 in skin at different developmental stages was examined with reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Along with fetal growth and development, the incidence rate of apoptosis was increased progressively in skin. In skin from early gestational fetus, bcl-2 was strongly expressed. This gene expression was progressively decreased with increment in gestational age. In children skin, the mRNA content of this gene was significantly reduced compared with fetal skin (P
4.Nasal anatomic abnormalities and bacterial infection in chronic rhino sinusitis.
Xinjun MENG ; Changping CAI ; Yuxing NI ; Shili WANG ; Jingyong SUN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2010;24(17):779-781
OBJECTIVE:
To study the relationship between nasal anatomic abnormalities and the bacteria infection status of maxillary sinus.
METHOD:
The anatomic abnormalities of 115 cases of maxillary sinuses were detected with the CT images and confirmed with chronic infection, which were divided into two groups: high anatomic abnormality group and low anatomic abnormality group. The sinusal contents were sent to bacteria culture, compare the bacteria infection rate and the distribution of bacteria between the two groups.
RESULT:
The bacteria positive rate of the high anatomic abnormality group and low anatomic abnormality group was 90.32% and 56.60% (P < 0.01) respectively. The contribution of gram-positive bacteria and gram-negative bacteria are 47.76% and 52.24% in high anatomic abnormality group, 62.16% and 37.84% in low anatomic abnormality group.
CONCLUSION
Nasal anatomic abnormalities can improve the bacteria infectious rate of maxillary sinuses. High anatomic abnormality may more induces gram-negative bacteria infections,while low anatomic abnormality may more induces gram-positive bacteria infections.
Adult
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Aged
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Aged, 80 and over
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Bacterial Infections
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etiology
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Chronic Disease
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Humans
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Middle Aged
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Nose
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abnormalities
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Sinusitis
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microbiology
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Young Adult
5.Exploration and thinking of online teaching of medical microbiology experiment
Mei QI ; Hong WANG ; Yabin ZHOU ; Juan LIU ; Wei TANG ; Yizhe CHENG ; Yundong SUN ; Wenjuan LI ; Shili LIU ; Weifang ZHANG
Chinese Journal of Medical Education Research 2023;22(4):528-531
Medical microbiology experiment is faced with many problems in online teaching. This study adopts the teaching mode of online live broadcast + operation video + virtual experiment, and make up the operation gap to some extent through operation video and virtual experiment. The mode of assessment is subjective thinking question (closely following the operation process) + experiment design + literature review (focusing on the key technology or new technology of clinical assessment that cannot be carried out due to the limitation of conditions in traditional experiments, such as mass spectrometry, fluorescence quantitative PCR, and G-test), and it is helpful to understand students' mastery of teaching objectives, and the ability of comprehensive application and innovative thinking. The student questionnaire shows that most students hold a positive attitude towards the online experimental teaching mode, and the quality of students' homework shows that most students have a good learning effect.
6.The in vitro tracing of miR-144/451 reveals the potential regulatory function of LINC01569 in erythropoiesis
Bingyi LIAO ; Wencui SUN ; Shili TANG ; Enxia HUANG ; Qingrong LIU ; Yuan XUE ; Yonggang ZHANG
Chinese Journal of Blood Transfusion 2024;37(5):516-523
【Objective】 Utilizing a specially engineered miR-144-GFP-H1 human embryonic stem cell (hESC) reporter line, this study leverages GFP fluorescence as an indicator of miR-144 expression to gauge the progression of erythropoiesis. The investigation is aimed at elucidating the potential roles of lncRNAs within the erythropoietic framework and conducting an initial assessment of their functional impact. 【Methods】 The miR-144/451-GFP-H1 cell line (hereafter referred to as 144-H1) was utilized for in vitro erythrocyte induction culture. The subpopulations of cells entering the erythropoiesis stage were characterized by the surface molecules CD71 and GPA. The GFP reporter gene of miR-144 served as a critical determinant to distinguish between GFP-positive cells (with a high propensity for erythropoiesis) and GFP-negative cells (with a low propensity for erythropoiesis). Transcriptome sequencing was performed on both groups to identify differentially expressed long non-coding RNAs (lncRNAs). LncRNA entries with potential for validation were selected for preliminary functional verification. The CRISPR/Cas9 gene editing technique was employed to design functional interference strategies for the targeted lncRNAs, obtaining 144-H1 cell lines with knocked-out function of the specific lncRNAs. These knockout cell lines, along with non-knockout 144-H1 cell lines, were used for parallel erythrocyte induction culture to identify differential nodes. This approach preliminarily verified their impact on erythropoiesis in an in vitro development model. 【Results】 1)The constructed 144-H1 cell line was capable of expressing GFP fluorescence upon entering the stage of in vitro erythrocyte induction, indicating the activation of miR-144/451. 2)Within the CD71, GPA double-positive group, significant differences in lncRNA expression were observed between the GFP-positive and GFP-negative subpopulations. 3) Gene editing strategies involving the deletion of sequence segments capable of effectively interfering with the function of multiple lncRNA entries were designed and verified for successful editing. In the knockout cell lines, parts of the lncRNA sequences were directly deleted. 4)In parallel validation experiments of erythrocyte induction culture, cell lines with LINC01569 knocked out exhibited significant differences in flow cytometric subpopulations and cell proliferation capabilities compared to the non-knockout cell lines: ①The knockout cell lines showed sustained high expression of GFP fluorescence. ②The proportion of the CD71-GPA double-positive group in the knockout cell lines continuously decreased during erythrocyte maturation. ③No significant expression of hemoglobin was observed in the knockout cell lines, lacking the characteristic red color. ④The cell proliferation capability of the knockout cell lines was significantly lower than that of the non-knockout cell lines (P<0.05). 【Conclusion】 The successful employment of the 144-H1 cell line facilitated an exploration into the potential functions of lncRNAs in erythropoiesis. This enables the design of more refined in vitro developmental experiments to enhance the precision in capturing lncRNA functions. Among the differentially expressed lncRNA entries, LINC01569 was preliminarily validated to play a regulatory role in erythropoiesis. The functional absence of LINC01569 severely impacts the normal differentiation and proliferation of erythrocytes. The specific regulatory mechanism of LINC01569 in erythropoiesis warrants further investigation and research.
7. Tea and Citrus maxima complex induces apoptosis of human liver cancer cells via PI3K/AKT/mTOR pathway in vitro
Shuai WEN ; Junxi CAO ; Zhigang LI ; Wenji ZHANG ; Ruohong CHEN ; Qiuhua LI ; Xingfei LAI ; Lingli SUN ; Shili SUN ; Ran AN ; Dongli LI ; Dongli LI
Chinese Herbal Medicines 2022;14(3):449-458
Objective: In this study, black tea and Citrus maxima (BT-CM), yellow tea and C. maxima (YT-CM), green tea and C. maxima (GT-CM) as subjects, the active ingredient content and antioxidant activity of three tea and C. maxima (T-CM) were analyzed. The effects of three T-CMs on apoptosis of liver cells in vitro and its mechanism were further explored. Methods: National standard method and HPLC were used for active ingredient analysis. MTT, cell flow cytometry and Western blot were used to analyze the effects of three T-CMs on cell proliferation, apoptosis, and its underlying molecular mechanism. Results: The content of tea polyphenols, free amino acids, ratio of polyphenols and amino acids, ester catechins, non-ester catechins and caffeine in YT-CM and GT-CM was significantly higher than that of BT-CM. The in vitro antioxidant capacity of YT-CM and GT-CM was also significantly stronger than that of BT-CM. Three T-CMs had the effects of inhibiting proliferation, arresting cell cycle and inducing apoptosis in HepG2 and Bel7402 cells, especially YT-CM and GT-CM. Western blot analysis showed three T-CMs activated PI3K/AKT/mTOR signaling pathway and regulated the expression levels of apoptosis-related proteins Bax, Bcl-2 and Caspase-3/9. YT-CM and GT-CM had better ability to change the signal pathway than BT-CM. Conclusion: In short, T-CMs, which combined different degrees of fermentation tea with C. maxima, were rich in nutrients and biologically active substances. T-CMs, especially YT-CM and GT-CM, are healthy drinks that help to prevent and treat liver cancer.