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.Analysis of respiratory syncytial virus detection outcomes of 973 cases with severe respiratory infection during 2016-2019, Yuhang district of Hangzhou
Shili SONG ; Mingqiang LI ; Yaping SUN
Chinese Journal of Preventive Medicine 2021;55(2):263-265
This study collected nasopharyngeal swab specimens from severe respiratory infection cases in First People′s Hospital of Yuhang District during 2016-2019. Real-time PCR was used to detect respiratory syncytial virus (RSV). Rate of RSV positive detection were analysised in different age groups and different months. A total of 973 nasopharyngeal swab specimens of severe respiratory infection cases were collected, and the total positive rate of nucleic acid test of RSV was 6.47%; The detection rate of nucleic acid in male is higher than that in female, with no statistical differences ( P=0.023). The positive rate of nucleic acid test was negatively correlated with age. The positive rate was 15.2% in the group aged 0-1 years and 12% in the group aged 1-2 years. There are obvious seasonal differences in the prevalence of RSV, human are easier to infect RSV in spring and winter.
6.Analysis of respiratory syncytial virus detection outcomes of 973 cases with severe respiratory infection during 2016-2019, Yuhang district of Hangzhou
Shili SONG ; Mingqiang LI ; Yaping SUN
Chinese Journal of Preventive Medicine 2021;55(2):263-265
This study collected nasopharyngeal swab specimens from severe respiratory infection cases in First People′s Hospital of Yuhang District during 2016-2019. Real-time PCR was used to detect respiratory syncytial virus (RSV). Rate of RSV positive detection were analysised in different age groups and different months. A total of 973 nasopharyngeal swab specimens of severe respiratory infection cases were collected, and the total positive rate of nucleic acid test of RSV was 6.47%; The detection rate of nucleic acid in male is higher than that in female, with no statistical differences ( P=0.023). The positive rate of nucleic acid test was negatively correlated with age. The positive rate was 15.2% in the group aged 0-1 years and 12% in the group aged 1-2 years. There are obvious seasonal differences in the prevalence of RSV, human are easier to infect RSV in spring and winter.
7.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.
8.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 determi-nant 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 ex-pressed long non-coding RNAs(lncRNAs).LncRNA entries with potential for validation were selected for preliminary func-tional 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 in-duction,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 de-leted.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 ex-pression 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 e-rythropoiesis.This enables the design of more refined in vitro developmental experiments to enhance the precision in captu-ring 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 prolif-eration of erythrocytes.The specific regulatory mechanism of LINC01569 in erythropoiesis warrants further investigation and research.
9.Activation of ALDH2 alleviates hypoxic pulmonary hypertension in mice by upregulating the SIRT1/PGC-1α signaling pathway
Lei WANG ; Fenlan BIAN ; Feiyang MA ; Shu FANG ; Zihan LING ; Mengran LIU ; Hongyan SUN ; Chengwen FU ; Shiyao NI ; Xiaoyang ZHAO ; Xinru FENG ; Zhengyu SUN ; Guoqing LU ; Pinfang KANG ; Shili WU
Journal of Southern Medical University 2024;44(10):1955-1964
Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.
10.Activation of ALDH2 alleviates hypoxic pulmonary hypertension in mice by upregulating the SIRT1/PGC-1α signaling pathway
Lei WANG ; Fenlan BIAN ; Feiyang MA ; Shu FANG ; Zihan LING ; Mengran LIU ; Hongyan SUN ; Chengwen FU ; Shiyao NI ; Xiaoyang ZHAO ; Xinru FENG ; Zhengyu SUN ; Guoqing LU ; Pinfang KANG ; Shili WU
Journal of Southern Medical University 2024;44(10):1955-1964
Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.