1.Dynamic observation on capillarization of liver sinusoidal endothelial cells induced by Echinococcus multilocularis infection
Renjie ZHANG ; Jun XIE ; Fanna WEI ; Xiaojin MO ; Peng SONG ; Yuchun CAI ; Yan LU ; Jiahui SUN ; Yan ZHOU ; Lin LIN ; Ting ZHANG ; Muxin CHEN
Chinese Journal of Schistosomiasis Control 2024;36(1):34-43
Objective To investigate the capillarization of liver sinusoidal endothelial cells (LSECs) and its association with hepatic fibrosis during the development of alveolar echinococcosis, so as to provide the basis for unraveling the mechanisms underlying the role of LSEC in the development and prognosis of hepatic injuries and hepatic fibrosis caused by alveolar echinococcosis. Methods Forty C57BL/6 mice at ages of 6 to 8 weeks were randomly divided into a control group and 1-, 2- and 4-week infection groups, of 10 mice in each group. Each mouse in the infection groups was intraperitoneally injected with 2 000 Echinococcus multilocularis protoscoleces, while each mouse in the control group was given an equal volume of phosphate-buffered saline using the same method. All mice were sacrificed 1, 2 and 4 weeks post-infection and mouse livers were collected. The pathological changes of livers were observed using hematoxylin-eosin (HE) staining, and hepatic fibrosis was evaluated through semi-quantitative analysis of Masson’s trichrome staining-positive areas. The activation of hepatic stellate cells (HSCs) and extracellular matrix (ECM) deposition were examined using immunohistochemical staining of α-smooth muscle actin (α-SMA) and collagen type I alpha 1 (COL1A1), and the fenestrations on the surface of LSECs were observed using scanning electron microscopy. Primary LSECs were isolated from mouse livers, and the mRNA expression of LSEC marker genes Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf was quantified using real-time fluorescence quantitative PCR (qPCR) assay. Results Destruction of local liver lobular structure was observed in mice 2 weeks post-infection with E. multilocularis protoscoleces, and hydatid cysts, which were surrounded by granulomatous tissues, were found in mouse livers 4 weeks post-infection. Semi-quantitative analysis of Masson’s trichrome staining showed a significant difference in the proportion of collagen fiber contents in mouse livers among the four groups (F = 26.060, P < 0.001), and a higher proportion of collagen fiber contents was detected in mouse livers in the 4-week infection group [(11.29 ± 2.58)%] than in the control group (P < 0.001). Immunohistochemical staining revealed activation of a few HSCs and ECM deposition in mouse livers 1 and 2 weeks post-infection, and abundant brown-yellow stained α-SMA and COL1A1 were deposited in the lesion areas in mouse livers 4 weeks post-infection, which spread to surrounding tissues. Semi-quantitative analysis revealed significant differences in α-SMA (F = 7.667, P < 0.05) and COL1A1 expression (F = 6.530, P < 0.05) in mouse levers among the four groups, with higher α-SMA [(7.13 ± 3.68)%] and COL1A1 expression [(13.18 ± 7.20)%] quantified in mouse livers in the 4-week infection group than in the control group (both P values < 0.05). Scanning electron microscopy revealed significant differences in the fenestration frequency (F = 37.730, P < 0.001) and porosity (F = 16.010, P < 0.001) on the surface of mouse LSECs among the four groups, and reduced fenestration frequency and porosity were observed in the 1-[(1.22 ± 0.48)/μm2 and [(3.05 ± 0.91)%] and 2-week infection groups [(3.47 ± 0.10)/μm2 and (7.57 ± 0.23)%] groups than in the control group (all P values < 0.001). There was a significant difference in the average fenestration diameter on the surface of mouse LSECs among the four groups (F = 15.330, P < 0.001), and larger average fenestration diameters were measured in the 1-[(180.80 ± 16.42) nm] and 2-week infection groups [(161.70 ± 3.85) nm] than in the control group (both P values < 0.05). In addition, there were significant differences among the four groups in terms of Stabilin-1 (F = 153.100, P < 0.001), Stabilin-2 (F = 57.010, P < 0.001), Ehd3 (F = 31.700, P < 0.001), CD209b (F = 177.400, P < 0.001), GATA4 (F = 17.740, P < 0.001), and Maf mRNA expression (F = 72.710, P < 0.001), and reduced mRNA expression of Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf genes was quantified in three infection groups than in the control group (all P values < 0.001). Conclusions E. multilocularis infections may induce capillarization of LSECs in mice, and result in a reduction in the expression of functional and phenotypic marker genes of LSECs, and capillarization of LSECs occurs earlier than activation of HSC and development of hepatic fibrosis.
2.Role of Akkermansia muciniphila in nonalcoholic fatty liver disease
Liting ZHENG ; Zhe WANG ; Yuchun CHEN ; Shanshan LIU ; Youcheng XIE ; Chuyi LI ; Xiaohui YU
Journal of Clinical Hepatology 2024;40(3):594-599
Nonalcoholic fatty liver disease (NAFLD) has become the most common chronic liver disease in the world, and it is also one of the main causes of liver cirrhosis and hepatocellular carcinoma, so it is particularly important to curb the development and progression of NAFLD in a timely manner. However, due to its complex pathogeneses, there are currently no effective methods for radical treatment. As a new generation of probiotics, Akkermansia muciniphila (Akk bacteria) can improve metabolic disorders of the body, and more and more studies have shown that Akk bacteria have a potential therapeutic effect on metabolic diseases, especially NAFLD. Therefore, this article briefly reviews the mechanism of action of Akk bacteria in NAFLD, in order to provide new ideas for improving the treatment of NAFLD and creating new therapies.
3.Overexpression of miR-320e inhibits inflammatory response of bronchial epithelial cells infected by respiratory syncytial virus
Zhenlang XU ; Xiangdong KUANG ; Jingchen XIE ; Yuchun QIN
Chinese Journal of Immunology 2024;40(12):2506-2512
Objective:To investigate the mechanism of overexpression of miR-320e in inhibiting inflammatory response of re-spiratory syncytial virus(RSV)infected bronchial epithelial cells.Methods:Human bronchial epithelial cells 16HBE were cultured in vitro and infected with RSV,and cells were divided into Con group,RSV group,RSV+miR-NC group,RSV+miR-320e group,RSV+miR-320e+vector group,RSV+miR-320e+TLR4 group.RT-qPCR was used to detect expression levels of miR-320e and TLR4 mRNA;MTT to detect cell proliferation changes;flow cytometry to detect cell apoptosis;Western blot was used to detect Bcl-2,Bax,TLR4,IκBα,p-IκBα,NF-κB and p-NF-κB protein expressions;ELISA to detect TNF-α,IL-6,IL-1β and IFN-α,IFN-β expres-sions;dual luciferase experiment to verify the tageting relationship between miR-320e and TLR4.Results:Compared with Con group,miR-320e expression level,survival rate,Bcl-2 and IκBα protein expressions were significantly reduced,apoptosis rate,Bax protein expression,TNF-α,IL-6,IL-1β and IFN-α,IFN-β expressions,TLR4 mRNA and protein expression,and p-IκBα protein expres-sion and p-NF-κB/NF-κB were increased significantly in RSV group.Compared with RSV+miR-NC group,miR-320e expression level,survival rate,IFN-α,IFN-β expressions,Bcl-2 and IκBα protein expressions were significantly increased,apoptosis rate,Bax pro-tein expression,TNF-α,IL-6,IL-1β expressions,TLR4 mRNA and protein expression,and p-IκBα protein expression and p-NF-κB/NF-κB in RSV+miR-320e group were significantly reduced.miR-320e targets and negatively regulates the expression of TLR4.Up-regulation of TLR4 can partially restore the effect of overexpression of miR-320e on apoptosis and inflammatory response of RSV-infected bronchial epithelial cells 16HBE.Conclusion:miR-320e inhibits 16HBE apoptosis and inflammation in RSV-infected bronchial epi-thelial cells by targeting and negatively regulating TLR4 expression.
4.Chromosomal microarray analysis of 17 patients with unbalanced reciprocal translocations.
Chonglan GAO ; Han KAN ; Xingyu LI ; Chengxiu XIE ; Yuchun PAN
Chinese Journal of Medical Genetics 2023;40(11):1360-1366
OBJECTIVE:
To retrospectively analyze the results of chromosomal microarray analysis (CMA) and parental origins of unbalanced translocations among 17 patients, so as to provide reference for their genetic counseling.
METHODS:
The results of CMA for 7 001 samples tested in Chengdu Women and Children's Central Hospital from January 2019 to January 2022 were retrospectively reviewed. Unbalanced reciprocal translocation was defined as two non-homologous chromosomes with lost and gained segments respectively or both with gained segments, and their parental origins were identified by parental chromosomal karyotyping and/or fluorescence in situ hybridization (FISH).
RESULTS:
In total 17 unbalanced translocations were identified. In three cases, two non-homologous chromosomes both had gained segments, which constituted a derivative chromosome, with the total number of chromosomes being 47. In the remaining 14 cases, there was a terminal deletion on one chromosome and a terminal duplication on the other, 10 of which were confirmed by karyotyping, with the total number of chromosomes being 46. In the derivative chromosome, the lost segment was replaced by a gained segment from another chromosome. Among 15 cases undergoing parental origin analysis, 12 had paternal or maternal chromosomal abnormalities, including 11 balanced translocations and 1 unbalanced translocation. The unbalanced gametes therefore may form through meiosis. In 3 cases, the parental chromosomes were normal, indicating a de novo origin.
CONCLUSION
Discovery of terminal duplication and deletion or gained segments on two non-homologous chromosomes by CMA is suggestive of parental balanced translocation, which can facilitate genetic counseling and assessment the recurrence risk for subsequent pregnancies.
Child
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Pregnancy
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Humans
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Female
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In Situ Hybridization, Fluorescence
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Retrospective Studies
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Translocation, Genetic
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Microarray Analysis
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Chromosomes
5.Study on the protective effect of rapamycin-treated dendritic cells on transfusion-related acute lung injury after infection
Li GAO ; Yiming YANG ; Rufeng XIE ; Yuchun GONG ; Juan SUN ; Xueyu JIANG ; Jie YANG
Chinese Journal of Blood Transfusion 2022;35(10):1005-1008
【Objective】 To investigate the viability of rapamycin-treated rapamycin-treated dendritic cells (DCs) in intervening transfusion-related acute lung injury (TRALI) after infection. 【Methods】 1)The TRALI mouse model was induced by lipopolysaccharide (LPS) combined with anti-H2Kd antibody. The mice anal temperature and the wet/dry ratio of lung, kidney, spleen and brain tissues were measured. 2) Mouse bone marrow-derived DC cells were induced in vitro and treated with rapamycin (10nM) for 24h. 3) Mice were injected with or without rapamycin or rapamycin-treated DC, then injected with LPS intraperitoneally one hour later, finally injected with anti-H2Kd antibody 24 hours later to induce the onset of TRALI. The death situation of the mice was observed and recorded. The condition of mice after the onset of TRALI was analyzed by mouse body temperature, lung wet-dry ratio, and pleural effusion weight and lung histopathological sections. 【Results】 By comparing the induction effects of anti-H2Kd antibody solutions with different concentrations and volumes, the mouse model induced by 0.1mg/kg LPS combined with 4.5 mg/kg anti-H2Kd antibody (infusion volume of 100μL) was selected as the TRALI mouse model for this study. After the onset of TRALI, the wet/dry ratio of the lungs could be significantly increased and the body temperature could be significantly reduced in the model mice. After the intervention of TRALI mice with DCs treated with rapamycin, the mortality rate was significantly reduced, and the lung tissue lesions of the mice were significantly improved, whose protection effect was better than that of the rapamycin-treated group. Compared with the TRALI incidence group, the weight of pleural effusion in the intervention group was significantly reduced (P<0.05), but there was no significant difference in lung wet/dry ratio and body temperature. 【Conclusion】 The combination of LPS and antibodies can effectively induce a stable and typical TRALI mouse model, suggesting that the presence of infectious inflammation and blood transfusion-related inflammatory substances are the decisive factor for the pathogenesis of TRALI. Meanwhile, DCs treated with rapamycin have a protective effect on post-infection transfusion-related acute lung injury, which is expected to be a potential cell therapy strategy to intervene in the exacerbation of TRALI.
6.A comparison between ACR TI-RADS and artificial intelligence TI-RADS regarding to diagnostic efficacy and ability to reduce unnecessary fine-needle aspiration cytology
Yuchun WANG ; Bin YANG ; Pengfei HUANG ; Yingdong XIE
Chinese Journal of Ultrasonography 2021;30(5):408-413
Objective:To compare the diagnostic efficacy and ability of Thyroid Imaging Reporting Data System version (TI-RADS) of American College of Radiology (ACR) and artificial intelligence(AI) TI-RADS in diagnosis of thyroid nodules.Methods:A retrospective analysis was done on 266 patients(276 nodules) proved by ultrasound-guided fine-needle aspiration cytology (US-FNAC) in General Hospital of Eastern Theater Command from January to December 2019. The ROC curve of the two TI-RADS versions was drawn and the area under the curve (AUC) was calculated and compared.Results:AUCs of ACR TI-RADS and AI TI-RADS were 0.747 and 0.853. The sensitivity, specificity, positive predictive value and negative predictive value (96.62%, 62.50%, 74.87%, 94.12%) of AI TI-RADS were higher than ACR TI-RADS (95.27%, 44.53%, 66.51%, 89.06%). AI TI-RADS was able to avoid more unnecessary FNAC (71.74%) than ACR TI-RADS (67.03%).Conclusions:Both ACR TI-RADS and AI TI-RADS have good performances for differential diagnosis of thyroid nodules. AI TI-RADS is a more simple scoring system with better overall diagnostic performance and ability to exclude unnecessary FNAC with high negative predictive value than ACR TI-RADS.
7.Comparison of MRI and CT for target volume delineation and dose coverage for partial breast irradiation in patients with breast cancer
Yuchun SONG ; Xin XIE ; Shunan CHE ; Guangyi SUN ; Yu TANG ; Jianghu ZHANG ; Jianyang WANG ; Hui FANG ; Bo CHEN ; Yongwen SONG ; Jing JIN ; Yueping LIU ; Shunan QI ; Yuan TANG ; Ningning LU ; Hao JING ; Yong YANG ; Ning LI ; Jing LI ; Shulian WANG ; Yexiong LI
Chinese Journal of Radiation Oncology 2021;30(3):244-248
Objective:To compare magnetic resonance imaging (MRI)-based and computed tomography (CT)-based target volume delineation and dose coverage in partial breast irradiation (PBI) for patients with breast cancer, aiming to explore the application value of MRI localization in PBI after breast-conserving surgery.Methods:Twenty-nine patients with early breast cancer underwent simulating CT and MRI scans in a supine position. The cavity visualization score (CVS) of tumor bed (TB) was evaluated. The TB, clinical target volume (CTV), planning target volume (PTV) were delineated on CT and MRI images, and then statistically compared. Conformity indices (CI) between CT- and MRI-defined target volumes were calculated. PBI treatment plan of 40 Gy in 10 fractions was designed based on PTV-CT, and the dose coverage for PTV-MRI was evaluated.Results:The CVS on CT and MRI images was 2.97±1.40 vs. 3.10±1.40( P=0.408). The volumes of TB, CTV, PTV on MRI were significantly larger than those on CT, (24.48±16.60) cm 3vs. (38.00±19.77) cm 3, (126.76±56.81) cm 3vs. (168.42±70.54) cm 3, (216.63±81.99) cm 3vs. (279.24±101.55) cm 3, respectively, whereas the increasing percentage of CTV and PTV were significantly smaller than those of TB. The CI between CT-based and MRI-based TB, CTV, PTV were 0.43±0.13, 0.66±0.11, 0.70±0.09( P<0.001), respectively. The median percentage of PTV-MRI receiving 40 Gy dose was 81.9%(62.3% to 92.4%), significantly lower than 95.6%(95.0%~97.5%) of PTV-CT. Conclusions:The CVS between CT and MRI is not significantly different, but the MRI-based TB, CTV, PTV are significantly larger than CT-based values. The PTV-MRI is of underdose if PBI treatment plan is designed for PTV-CT. As a supplement of CT scan, MRI can enhance the accuracy of TB delineation after breast-onserving surgery.
8.Genetic analysis and prenatal diagnosis for ten families affected with tuberous sclerosis complex.
Yuchun PAN ; Weiqing WU ; Caiqun LUO ; Jiansheng XIE ; Zhiyong XU ; Qian GENG ; Ying HAO
Chinese Journal of Medical Genetics 2018;35(1):18-22
OBJECTIVE To provide prenatal diagnosis for families affected with tuberous sclerosis complex and explore the correlation between phenotype and genotype. METHODS For probands from 10 families, all exons and splicing regions of the TSC1 and TSC2 genes were analyzed with high throughput DNA sequencing. Suspected mutations were verified by Sanger sequencing. RESULTS All probands were found to have mutations, which included 1 case with TSC1 mutation and 9 cases with TSC2 mutations (missense mutations in 6, nonsense mutations in 2, and frameshifting mutation in 1 case). Prenatal diagnosis was provided for 9 cases, and 1 fetus was found to carry a mutation. Genetic analysis has identified a novel pathogenic mutation (TSC2 c.2415-2416 ins GT). CONCLUSION Identification of pathological mutations for tuberous sclerosis complex can facilitate genetic counseling and prenatal diagnosis for the affected families.
9.Analysis of 12 patients with novel mutations of Dystrophin gene.
Xiaoxin XU ; Yang LIU ; Yuchun PAN ; Zhiyong XU ; Qin WANG ; Jiangsheng XIE
Chinese Journal of Medical Genetics 2017;34(6):802-805
OBJECTIVETo study the characteristics, location, and amino acid changes of novel mutations of the Dystrophin gene.
METHODSTwelve patients in whom no deletion or duplication of the Dystrophin gene was detected were analyzed with next-generation sequencing. Fifty healthy adult males were recruited as the controls.
RESULTSAll patients were detected with mutations of the Dystrophin gene, which included c.33C>G, c.583C>T, c.1333C>T, c.2593C>T, c.5731A>T, c.7288G>T, c.2803+1G>T, c.10034G>A, c.4289A>G, c.1905_906delAG, c.5017delC, c.5768_5771delAAGA, and c.6261_6262insA. No similar mutations were found among the controls.
CONCLUSIONOur data has enriched the mutation spectrum of the Dystrophin gene and may provide an important basis for genetic diagnosis.
Child ; Child, Preschool ; Dystrophin ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Male ; Mutation
10.Gao Yuchun's experience of facial paralysis treatment.
Yanjun WANG ; Linhua CUI ; Jun YUAN ; Li HE ; Zhanqing XIE ; Weihua XUE ; Mei LI ; Zhenwei ZHANG ; Yuchun GAO ; Suobin KANG
Chinese Acupuncture & Moxibustion 2015;35(5):479-482
To introduce professor Gao Yuchun's clinical experience and treating characteristics of facial paralysis treated with acupuncture and moxibustion. Professor Gao pays attention to yangming when he selects acupoints for clinical syndrome, and directs acupoints selection based on syndrome differentiation in different levels of jingjin, meridians and zangfu; he praises opposing needling technique and reinforcing the deficiency and reducing the excess highly; the acupuncture manipulation is gentle,shallow and slow for reducing the healthy side and reinforcing the affected side, and through losing its excess to complement its deficiency; besides, he stresses needle retaining time and distinguishes reinforcing and reducing. Facial paralysis is treated with key factors such as acupoints selecting based on yangming, acupuncture manipulation, needle retaining time, etc. And the spleen and stomach is fine and good at transportation and transformation; the meridians is harmonious; the qi and blood is smooth. The clinical efficacy is enhanced finally.
Acupuncture Points
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Acupuncture Therapy
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history
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methods
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Aged, 80 and over
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China
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Facial Paralysis
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history
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therapy
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History, 20th Century
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History, 21st Century
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Humans

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