1.Construction of a mouse model for alveolar type Ⅱ epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system
Kun YANG ; Rong ZHANG ; Yue WU ; Xiaoping LEI ; Yunchuan SHEN ; Lan KANG ; Wenbin DONG
Chinese Journal of Tissue Engineering Research 2025;29(14):2943-2950
BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.
2.Significance of respiratory event duration in evaluating nocturnal hypoxemia and disease severity in adults with obstructive sleep apnea
Yiming WANG ; Chengcheng LIU ; Wenbin GUO ; Lin SUN ; Jinping ZENG ; Zhuqi CHEN ; Huijun YUE ; Wenbin LEI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(8):868-874
Objective:To analyze the correlation between respiratory event duration and nocturnal oxygen saturation (SpO 2) in adults with obstructive sleep apnea (OSA), and to explore its significance in assessing nocturnal hypoxemia and OSA severity. Methods:A prospective study was conducted on adult OSA patients diagnosed via overnight standard polysomnography (PSG) at the Department of Otolaryngology, First Affiliated Hospital of Sun Yat-sen University from June 2019 to December 2023. Data collected included demographic information, PSG reports, scale scores, and comorbidities. Patients were first stratified by apnea-hypopnea index (AHI) severity. Relationships between respiratory event duration parameters,including total apnea-hypopnea time (TAHT), percentage of total sleep time with apnea-hypopnea (AHT%), total apnea time (TAT), total hypopnea time (THT), and mean apnea-hypopnea time (MAHT), and nocturnal SpO? parameters, including average SpO? (aSpO?), minimal SpO? (mSpO?), mean oxygen desaturation (MOD), and percentage of total sleep time with SpO?<90% (CT90), were analyzed. Patients were then divided into two groups based on the median MAHT (27.6 s) for SpO? comparison. Finally, severe OSA patients were further subclassified using an AHI inflection point (50 events/h) identified via scatter plot analysis to compare nocturnal SpO?. Statistical analysis was performed using SPSS 27.0.Results:Among the 250 study subjects, there were 201 males and 49 females, with ages ranging from 18 to 76 years (mean age: 41.6 ± 11.9 years).TAHT, AHT%, and TAT in OSA patients demonstrated significant negative correlations with aSpO?( r=-0.698, -0.718, -0.646)and mSpO?( r=-0.746, -0.746, -0.748), while showing positive correlations with MOD ( r=0.783, 0.791, 0.823)and CT90 ( r=0.868, 0.866, 0.852), P<0.05. When stratified by MAHT median ( M=27.6 s), the "long-event" subgroup ( n=125) displayed significantly lower mSpO 2 and higher MOD and CT90 compared to the "short-event" subgroup ( n=125), Z=-3.319, 3.288, 2.242; P<0.05. No significant difference in aSpO 2 was observed ( P>0.05). Subgrouping severe OSA patients at AHI=50 events/hour revealed significant differences in aSpO 2, mSpO 2, MOD, and CT90 between groups ( Z=-5.011, -4.787, 5.142, 6.117, P<0.05). Conclusions:TAHT, AHT%, and TAT significantly correlate with nocturnal SpO? parameters in OSA patients and can supplement AHI in assessing OSA severity. MAHT independently reflects nocturnal oxygenation status beyond AHI.
3.Pathogenesis and intervention strategies of abdominal aortic aneurysm:an update
Yue CHAI ; Hongming SHAO ; Wenbin WU ; Ting HAN ; Pei WANG ; Chunlin ZHUANG
Academic Journal of Naval Medical University 2025;46(6):790-797
Abdominal aortic aneurysm(AAA)is a degenerative vascular disease occurring in the lower segment of the aortic diaphragm,mainly manifested by irreversible dilatation of the entire artery,preferably in the elderly population.The pathogenesis of AAA is complex and involves multiple factors,with genetic variations and immune imbalances playing important roles.Its pathological changes mainly include inflammatory cell infiltration,degradation of stromal elastin,and smooth muscle cell death.Rupture of AAA is the most dangerous complication,with a high mortality.Surgery remains the only effective intervention,but carries certain risks and postoperative complications.Early intervention for small abdominal aortic aneurysms to slow down aneurysm expansion and achieve long-term survival is currently a focus of drug and technology research.This article reviews the pathogenesis of AAA and its intervention strategies,and summarizes the research on existing drugs and the use of new targets and technologies,so as to provide insights for better understanding and treatment of AAA.
4.Feasibility and safety of mechanical thrombectomy in the endovascular recanalization of non-acute symptomatic long-segment internal carotid artery occlusion
Lanqi LI ; Chao LI ; Mingchao SHI ; Dajiang XING ; Jie ZHOU ; Feixue YUE ; Kangjia SONG ; Shouchun WANG ; Wenbin ZHANG
Chinese Journal of Cerebrovascular Diseases 2025;22(10):665-676
Objective To explore the safety and feasibility of mechanical thrombectomy as the first-line endovascular strategy in patients with non-acute symptomatic long-segment internal carotid artery occlusion(ICAO)undergoing revascularization.Methods This study retrospectively and consecutively enrolled non-acute symptomatic long-segment ICAO patients treated in the Department of Neurology,First Hospital of Jilin University,between January 2019 and August 2023,with mechanical thrombectomy as the preferred endovascular modality.Baseline and clinical data were collected,including sex,age,stroke-related risk factors(hypertension,diabetes,dyslipidemia,coronary artery disease,prior stroke,smoking and alcohol use history),admission National Institutes of Health stroke scale(NIHSS)score,pre-operative modified Rankin scale(mRS)score,time from last symptom onset to femoral puncture,time from imaging confirmation to femoral puncture,high-resolution MRI,right-sided ICAO,stump morphology(absent,tapered,flat/blunt,irregular),distal backfilling patterns(above ophthalmic segment,cavernous/clinoid segment,below cavernous segment),pathogenesis(atherosclerosis,dissection),types of anesthesia(local,general),procedure time(time frame from femoral puncture to recanalization or final angiography),site of the original occlusion in successfully recanalized cases,surgical techniques(aspiration+balloon angioplasty,aspiration+balloon angioplasty+stent-retriever thrombectomy,aspiration+balloon angioplasty+stent placement,aspiration+balloon angioplasty+stent-retriever thrombectomy+stent placement),stent placement(yes/no),number of stents implanted,and number of cases with retrieved thrombus,observed indicators.Observed indicators including ratio of technical successful recanalization(immediately post-procedure most severely stenosed site stenosis rate<50%,expanded thrombolysis in cerebral infarction[eTICI]grade≥2c),intraoperative complications(distal embolization,symptomatic intracranial hemorrhage,arterial perforation)rate,perioperative mortality rate,30-day stroke recurrence,and 90-day mRS score.Compare the baseline data,clinical data and observational indicators of the patients with successful and unsuccessful recanalization.Base on the original occlusion site,successfully recanalized patients were subclassified into isolated extracranial,isolated intracranial,and tandem lesions patients,and their baseline characteristics and observation indicators were compared.Results(1)A total of 65 patients were enrolled(57 men,8 women;age 39-80 years;median 59[52,65]years)in this study.Technical success was achieved in 52cases(80%).Perioperative complications occurred in 4 patients(6.2%),with 3 distal embolization cases(4.6%),1(1.5%)developed symptomatic intracranial hemorrhage,and no arterial perforations were observed.There was no perioperative mortality.The 30-day stroke recurrence rate was 7.7%(5/65).90-day mRS scores ranged from 0 to 4,with a median of 1.0(0.0,1.5).(2)Baseline and clinical characteristics as well as outcome indicators did not differ significantly between patients with successful versus unsuccessful recanalization in the cohort undergoing mechanical thrombectomy for non-acute symptomatic long-segment intracranial carotid artery occlusion(all P>0.05).(3)Among successfully recanalized patients,17(32.7%)had isolated extracranial lesions,18(34.6%)had isolated intracranial lesions,and 17(32.7%)had tandem lesions.All cases in the extracranial lesions group had original lesion site at the origin of internal carotid artery(C1,17/17).The intracranial group most often had orginal lesion sites at the C4 segment(9/18),whereas tandem lesions predominantly involved C1 plus C4-C5(16/17).Among the three groups,patients with isolated intracranial lesions were younger(57[48,61]years vs.60[52,64],63[58,69]years,P=0.050),and had a lower proportion of right-sided ICAO(4/18 vs.11/17 vs.11/17,P=0.032),while patients with tandem lesions required a greater number of stents(2.0[1.0,2.0]vs.1.0[1.0,1.5],1.0[0.8,2.0],P=0.013).Significant differences were observed in the proportion of patients with retrieved thrombus decreased progressively from patients with isolated extracranial,isolated intracranial to tandem lesions(17/17 vs.17/18 vs.12/17,P=0.024).No significant differences were observed among lesion-site groups with respect to medical history,stump morphology,distal retrograde flow,procedural technique,procedure duration,anesthesia method,or outcome indicators(all P>0.05).Conclusions This study suggested that utilizing mechanical thrombectomy as the first-line endovascular therapy for non-acute symptomatic long-segment ICAO is safe and feasible.The original occlusive sites of non-acute symptomatic long-segment ICAO predominantly involve the cervical origin and the cavernous segment of the internal carotid artery.The conclusions of this study require further validation.
5.Significance of respiratory event duration in evaluating nocturnal hypoxemia and disease severity in adults with obstructive sleep apnea
Yiming WANG ; Chengcheng LIU ; Wenbin GUO ; Lin SUN ; Jinping ZENG ; Zhuqi CHEN ; Huijun YUE ; Wenbin LEI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(8):868-874
Objective:To analyze the correlation between respiratory event duration and nocturnal oxygen saturation (SpO 2) in adults with obstructive sleep apnea (OSA), and to explore its significance in assessing nocturnal hypoxemia and OSA severity. Methods:A prospective study was conducted on adult OSA patients diagnosed via overnight standard polysomnography (PSG) at the Department of Otolaryngology, First Affiliated Hospital of Sun Yat-sen University from June 2019 to December 2023. Data collected included demographic information, PSG reports, scale scores, and comorbidities. Patients were first stratified by apnea-hypopnea index (AHI) severity. Relationships between respiratory event duration parameters,including total apnea-hypopnea time (TAHT), percentage of total sleep time with apnea-hypopnea (AHT%), total apnea time (TAT), total hypopnea time (THT), and mean apnea-hypopnea time (MAHT), and nocturnal SpO? parameters, including average SpO? (aSpO?), minimal SpO? (mSpO?), mean oxygen desaturation (MOD), and percentage of total sleep time with SpO?<90% (CT90), were analyzed. Patients were then divided into two groups based on the median MAHT (27.6 s) for SpO? comparison. Finally, severe OSA patients were further subclassified using an AHI inflection point (50 events/h) identified via scatter plot analysis to compare nocturnal SpO?. Statistical analysis was performed using SPSS 27.0.Results:Among the 250 study subjects, there were 201 males and 49 females, with ages ranging from 18 to 76 years (mean age: 41.6 ± 11.9 years).TAHT, AHT%, and TAT in OSA patients demonstrated significant negative correlations with aSpO?( r=-0.698, -0.718, -0.646)and mSpO?( r=-0.746, -0.746, -0.748), while showing positive correlations with MOD ( r=0.783, 0.791, 0.823)and CT90 ( r=0.868, 0.866, 0.852), P<0.05. When stratified by MAHT median ( M=27.6 s), the "long-event" subgroup ( n=125) displayed significantly lower mSpO 2 and higher MOD and CT90 compared to the "short-event" subgroup ( n=125), Z=-3.319, 3.288, 2.242; P<0.05. No significant difference in aSpO 2 was observed ( P>0.05). Subgrouping severe OSA patients at AHI=50 events/hour revealed significant differences in aSpO 2, mSpO 2, MOD, and CT90 between groups ( Z=-5.011, -4.787, 5.142, 6.117, P<0.05). Conclusions:TAHT, AHT%, and TAT significantly correlate with nocturnal SpO? parameters in OSA patients and can supplement AHI in assessing OSA severity. MAHT independently reflects nocturnal oxygenation status beyond AHI.
6.Feasibility and safety of mechanical thrombectomy in the endovascular recanalization of non-acute symptomatic long-segment internal carotid artery occlusion
Lanqi LI ; Chao LI ; Mingchao SHI ; Dajiang XING ; Jie ZHOU ; Feixue YUE ; Kangjia SONG ; Shouchun WANG ; Wenbin ZHANG
Chinese Journal of Cerebrovascular Diseases 2025;22(10):665-676
Objective To explore the safety and feasibility of mechanical thrombectomy as the first-line endovascular strategy in patients with non-acute symptomatic long-segment internal carotid artery occlusion(ICAO)undergoing revascularization.Methods This study retrospectively and consecutively enrolled non-acute symptomatic long-segment ICAO patients treated in the Department of Neurology,First Hospital of Jilin University,between January 2019 and August 2023,with mechanical thrombectomy as the preferred endovascular modality.Baseline and clinical data were collected,including sex,age,stroke-related risk factors(hypertension,diabetes,dyslipidemia,coronary artery disease,prior stroke,smoking and alcohol use history),admission National Institutes of Health stroke scale(NIHSS)score,pre-operative modified Rankin scale(mRS)score,time from last symptom onset to femoral puncture,time from imaging confirmation to femoral puncture,high-resolution MRI,right-sided ICAO,stump morphology(absent,tapered,flat/blunt,irregular),distal backfilling patterns(above ophthalmic segment,cavernous/clinoid segment,below cavernous segment),pathogenesis(atherosclerosis,dissection),types of anesthesia(local,general),procedure time(time frame from femoral puncture to recanalization or final angiography),site of the original occlusion in successfully recanalized cases,surgical techniques(aspiration+balloon angioplasty,aspiration+balloon angioplasty+stent-retriever thrombectomy,aspiration+balloon angioplasty+stent placement,aspiration+balloon angioplasty+stent-retriever thrombectomy+stent placement),stent placement(yes/no),number of stents implanted,and number of cases with retrieved thrombus,observed indicators.Observed indicators including ratio of technical successful recanalization(immediately post-procedure most severely stenosed site stenosis rate<50%,expanded thrombolysis in cerebral infarction[eTICI]grade≥2c),intraoperative complications(distal embolization,symptomatic intracranial hemorrhage,arterial perforation)rate,perioperative mortality rate,30-day stroke recurrence,and 90-day mRS score.Compare the baseline data,clinical data and observational indicators of the patients with successful and unsuccessful recanalization.Base on the original occlusion site,successfully recanalized patients were subclassified into isolated extracranial,isolated intracranial,and tandem lesions patients,and their baseline characteristics and observation indicators were compared.Results(1)A total of 65 patients were enrolled(57 men,8 women;age 39-80 years;median 59[52,65]years)in this study.Technical success was achieved in 52cases(80%).Perioperative complications occurred in 4 patients(6.2%),with 3 distal embolization cases(4.6%),1(1.5%)developed symptomatic intracranial hemorrhage,and no arterial perforations were observed.There was no perioperative mortality.The 30-day stroke recurrence rate was 7.7%(5/65).90-day mRS scores ranged from 0 to 4,with a median of 1.0(0.0,1.5).(2)Baseline and clinical characteristics as well as outcome indicators did not differ significantly between patients with successful versus unsuccessful recanalization in the cohort undergoing mechanical thrombectomy for non-acute symptomatic long-segment intracranial carotid artery occlusion(all P>0.05).(3)Among successfully recanalized patients,17(32.7%)had isolated extracranial lesions,18(34.6%)had isolated intracranial lesions,and 17(32.7%)had tandem lesions.All cases in the extracranial lesions group had original lesion site at the origin of internal carotid artery(C1,17/17).The intracranial group most often had orginal lesion sites at the C4 segment(9/18),whereas tandem lesions predominantly involved C1 plus C4-C5(16/17).Among the three groups,patients with isolated intracranial lesions were younger(57[48,61]years vs.60[52,64],63[58,69]years,P=0.050),and had a lower proportion of right-sided ICAO(4/18 vs.11/17 vs.11/17,P=0.032),while patients with tandem lesions required a greater number of stents(2.0[1.0,2.0]vs.1.0[1.0,1.5],1.0[0.8,2.0],P=0.013).Significant differences were observed in the proportion of patients with retrieved thrombus decreased progressively from patients with isolated extracranial,isolated intracranial to tandem lesions(17/17 vs.17/18 vs.12/17,P=0.024).No significant differences were observed among lesion-site groups with respect to medical history,stump morphology,distal retrograde flow,procedural technique,procedure duration,anesthesia method,or outcome indicators(all P>0.05).Conclusions This study suggested that utilizing mechanical thrombectomy as the first-line endovascular therapy for non-acute symptomatic long-segment ICAO is safe and feasible.The original occlusive sites of non-acute symptomatic long-segment ICAO predominantly involve the cervical origin and the cavernous segment of the internal carotid artery.The conclusions of this study require further validation.
7.Construction of a mouse model for alveolar type Ⅱ epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system
Kun YANG ; Rong ZHANG ; Yue WU ; Xiaoping LEI ; Yunchuan SHEN ; Lan KANG ; Wenbin DONG
Chinese Journal of Tissue Engineering Research 2025;29(14):2943-2950
BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.
8.To explore the mechanism of Polygonatum and Astragalus compound in inhibiting lung adenocarcinoma based on APELIN-PGC1α-UCP1 signaling pathway
Zongcan WANG ; Tiansheng ZHENG ; Mengling WEI ; Wenbin ZHUANG ; Ming LI ; Fei WANG ; Liduo YUE ; Lihong FAN
Tumor 2024;44(2):180-194
Objective:To investigate the mechanism of polygonatum and astragalus compound(PA)in inhibiting the progression of lung adenocarcinoma. Methods:CCK-8 assay was used to assess the inhibitory rate of proliferation in A549 and H1299 cells treated with PA at different concentrations and to calculate the half maximal inhibitory concentration(IC50).C57BL/6 mice(KRASG12D/+;TP53flox/flox)were treated with adenovirus carrying Cre enzyme via nasal inhalation to establish a mouse model of primary lung adenocarcinoma.The model mice were fed with PA-containing diet to directly observe the effect of PA on the lung adenocarcinoma tissue.Immunohistochemical staining was used to examine the pathological status of the lung tissue.Bioinformatics analysis indicated that PA affects the progression of lung adenocarcinoma through the apelin-peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC1α)-mitochondrial brown fat uncoupling protein 1(UCP1).Real-time quantitative PCR and Western blotting analysis were used to study the effect of PA on the mRNA and protein expression levels of apelin-PGC1α-UCP1 signaling pathway related genes.An ATP detection kit and flow cytometry were used to evaluate the effect of PA on the ATP and mitochondrial ROS production,respectively,in A549 and H1299 cells.siUCP1 was used to silent the expression of UCP1 while Z160 was used to induce UCP1 overexpression in A549 and H1299 cells,and the changes in ATP and mitochondrial ROS production were examined to further investigate whether PA acts on apelin-PGC1α-UCP1 signaling pathway to affect the progression of lung adenocarcinoma. Results:PA could obviously inhibit the proliferation of A549 and H1299 cells with the IC50 values of 10.66 mg/mL for A549 cells and 9.66 mg/mL for H1299 cells.In the mouse primary lung adenocarcinoma model,PA could effectively inhibit the growth of tumor,downregulate apelin-PGC1α-UCP1 signaling pathway and inhibit the expression of lung adenocarcinoma-promoting gene UCP1.In A549 and H1299 cells,PA could significantly inhibit the expression of apelin,PGC1α and UCP1(P<0.05),promote the production of ATP(P<0.000 1)and ROS,restore mitochondrial oxidative phosphorylation,and inhibit aerobic glycolysis(P<0.01).UCP1 silencing could increase the production of ATP(P<0.01)and mitochondrial ROS and decrease the expression of key glycolysis enzymes hexokinase 2(HK2)and pyruvate kinase isozyme type M2(PKM2)(P<0.05).Increasing the expression of UCP1 could reduce the ATP production(P<0.01)and mitochondrial ROS generation in cells while increase the expression of HK2 and PKM2(P<0.05).Treating cells with PA and Z160 simultaneously(PA+Z160)could reverse the inhibitory effect of PA on the ATP production and glycolysis of tumor cells(P<0.05). Conclusion:PA can downregulate the apelin-PGC1α-UCP1 signaling pathway,inhibit mitochondrial uncoupling,restore mitochondrial oxidative phosphorylation,inhibit aerobic glycolysis,reverse the Warburg effect,and thus inhibit lung adenocarcinoma progression.
9.CRISPR-assisted transcription activation by phase-separation proteins.
Jiaqi LIU ; Yuxi CHEN ; Baoting NONG ; Xiao LUO ; Kaixin CUI ; Zhan LI ; Pengfei ZHANG ; Wenqiong TAN ; Yue YANG ; Wenbin MA ; Puping LIANG ; Zhou SONGYANG
Protein & Cell 2023;14(12):874-887
The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been widely used for genome engineering and transcriptional regulation in many different organisms. Current CRISPR-activation (CRISPRa) platforms often require multiple components because of inefficient transcriptional activation. Here, we fused different phase-separation proteins to dCas9-VPR (dCas9-VP64-P65-RTA) and observed robust increases in transcriptional activation efficiency. Notably, human NUP98 (nucleoporin 98) and FUS (fused in sarcoma) IDR domains were best at enhancing dCas9-VPR activity, with dCas9-VPR-FUS IDR (VPRF) outperforming the other CRISPRa systems tested in this study in both activation efficiency and system simplicity. dCas9-VPRF overcomes the target strand bias and widens gRNA designing windows without affecting the off-target effect of dCas9-VPR. These findings demonstrate the feasibility of using phase-separation proteins to assist in the regulation of gene expression and support the broad appeal of the dCas9-VPRF system in basic and clinical applications.
Humans
;
Transcriptional Activation
;
RNA, Guide, CRISPR-Cas Systems
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Gene Expression Regulation
;
CRISPR-Cas Systems/genetics*
10.Machine and deep learning-based clinical characteristics and laboratory markers for the prediction of sarcopenia.
He ZHANG ; Mengting YIN ; Qianhui LIU ; Fei DING ; Lisha HOU ; Yiping DENG ; Tao CUI ; Yixian HAN ; Weiguang PANG ; Wenbin YE ; Jirong YUE ; Yong HE
Chinese Medical Journal 2023;136(8):967-973
BACKGROUND:
Sarcopenia is an age-related progressive skeletal muscle disorder involving the loss of muscle mass or strength and physiological function. Efficient and precise AI algorithms may play a significant role in the diagnosis of sarcopenia. In this study, we aimed to develop a machine learning model for sarcopenia diagnosis using clinical characteristics and laboratory indicators of aging cohorts.
METHODS:
We developed models of sarcopenia using the baseline data from the West China Health and Aging Trend (WCHAT) study. For external validation, we used the Xiamen Aging Trend (XMAT) cohort. We compared the support vector machine (SVM), random forest (RF), eXtreme Gradient Boosting (XGB), and Wide and Deep (W&D) models. The area under the receiver operating curve (AUC) and accuracy (ACC) were used to evaluate the diagnostic efficiency of the models.
RESULTS:
The WCHAT cohort, which included a total of 4057 participants for the training and testing datasets, and the XMAT cohort, which consisted of 553 participants for the external validation dataset, were enrolled in this study. Among the four models, W&D had the best performance (AUC = 0.916 ± 0.006, ACC = 0.882 ± 0.006), followed by SVM (AUC =0.907 ± 0.004, ACC = 0.877 ± 0.006), XGB (AUC = 0.877 ± 0.005, ACC = 0.868 ± 0.005), and RF (AUC = 0.843 ± 0.031, ACC = 0.836 ± 0.024) in the training dataset. Meanwhile, in the testing dataset, the diagnostic efficiency of the models from large to small was W&D (AUC = 0.881, ACC = 0.862), XGB (AUC = 0.858, ACC = 0.861), RF (AUC = 0.843, ACC = 0.836), and SVM (AUC = 0.829, ACC = 0.857). In the external validation dataset, the performance of W&D (AUC = 0.970, ACC = 0.911) was the best among the four models, followed by RF (AUC = 0.830, ACC = 0.769), SVM (AUC = 0.766, ACC = 0.738), and XGB (AUC = 0.722, ACC = 0.749).
CONCLUSIONS:
The W&D model not only had excellent diagnostic performance for sarcopenia but also showed good economic efficiency and timeliness. It could be widely used in primary health care institutions or developing areas with an aging population.
TRIAL REGISTRATION
Chictr.org, ChiCTR 1800018895.
Humans
;
Aged
;
Sarcopenia/diagnosis*
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Deep Learning
;
Aging
;
Algorithms
;
Biomarkers

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