3.Expression and clinical significance of plasma exosomal miR-34-5p and miR-142-3p in systemic sclerosis.
Wen Gen LI ; Xiao Dong GU ; Rui Qiang WENG ; Su Dong LIU ; Chao CHEN
Journal of Peking University(Health Sciences) 2023;55(6):1022-1027
OBJECTIVE:
To detect the expression of plasma exosomal microRNA (miRNA) in systemic sclerosis (SSc), and to investigate its clinical significance.
METHODS:
A total of 20 patients who were initially diagnosed with SSc and did not receive medication in Department of Rheumatology and Immunology of Meizhou People' s Hospital from January 2020 to January 2022 were recruited, as well as 15 healthy individuals whose gender and age matched with those of the SSc patients. Plasma exosomes were isolated using ultracentrifugation method. The expression levels of exosomal miR-34-5p, miR-92-3p and miR-142-3p were detected by quantative real-time polymerase chain reaction (qRT-PCR). Correlations between the expression levels of exosomal miRNAs and clinical characteristic were analyzed by Spearman's rank correlation coefficient test.
RESULTS:
The mean age of 20 patients with SSc was (52.6±12.6) years, including 7 males and 13 females. Among the 20 SSc patients, 13 cases were diagnosed as limited cutaneous systemic sclerosis (lcSSc) and 7 cases were diagnosed as diffuse cutaneous systemic sclerosis (dcSSc) according to the extent of skin involvement. According to the findings of high resolution chest CT, 7 of 20 SSc patients were diagnosed with interstitial lung disease (ILD) and 13 SSc patients were diagnosed with non-ILD. The expression levels of exosomal miR-34-5p, miR-92-3p and miR-142-3p were significantly elevated in the SSc patients compared with those in the healthy controls group (P=0.003, P=0.000 1, and P=0.016, respectively). Compared with the SSc patients without ILD, the expression levels of miR-34-5p and miR-142-3p were significantly lower in the SSc patients with ILD (P=0.037 and P=0.015, respectively). The expression levels of exosomal miR-34-5p and miR-142-3p showed negative correlation with ILD (r=-0.48, P=0.031 and r=-0.55, P=0.011, respectively), and arthritis (r=-0.46, P=0.040 and r=-0.48, P=0.032, respectively). The expression levels of exosomal miR-142-3p showed a negative correlation with erythrocyte sedimentation rate (ESR) (r=-0.55, P=0.012).
CONCLUSION
Plasma exosomal miR-34-5p, miR-92-3p and miR-142-3p were dysregulated in SSc. The dyregulation of exosomal miR-34-5p and miR-142-3p showed correlation with SSc associated ILD (SSc-ILD).
Male
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Female
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Humans
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Young Adult
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Adult
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Clinical Relevance
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MicroRNAs/genetics*
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Scleroderma, Systemic/genetics*
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Lung Diseases, Interstitial
4.Study on effect and mechanism of HIF -1 α silencing combined with methylselenenic acid on proliferation and apoptosis of cervical cancer cells
Zhenhao Li ; Yaofeng Hou ; Ling Li ; Xuanyu Huang ; Wanyu Liu ; Aixia Zhang ; Nan Wang
Acta Universitatis Medicinalis Anhui 2022;57(12):1954-1959
Objective :
To investigate the influence and molecular mechanism of hypoxia-inducing factor-1 α( HIF- 1 α) gene silencing combined with methyl selenenic acid (MSA) on cervical cancer cell proliferation,apoptosis and cell migration.
Methods :
HeLa cells were transfected with HIF-1 interference RNA and negative control RNA.Af- ter transfection for 48 h,cells were stimulated with MSA for 24 h,and cell proliferation was determined by CCK-8 assay and colony formation.Apoptosis was determined by flow cytometry combined with Annexin V-FITC / PI.The expression levels of HIF-1α , Bcl-2 ,and E-cadherin were detected by Western blot assay. Cell migration ability was determined by Transwell assay. RNA-seq analysis was used to investigate the differentially expressed genes and differential signaling pathways.
Results :
Compared with the control group,interfering with HIF-1α combined with MSA significantly inhibited cell proliferation (P <0.01) .Flow cytometry results showed that the combined drug group significantly induced apoptosis.Transwell results showed that interfering with HIF-1α combined with MSA
inhibited HeLa cell migration.Compared with the control group,interfering with HIF-1α combined with MSA down- regulated the expression of Bcl-2 and up-regulated the expression of E-cadherin. RNA-sequencing combined with signal pathway enrichment results showed that the expression of apoptotic signal pathway and downstream genes was inhibited.
Conclusion
HIF-1α gene silencing combined with MSA can synergically inhibit the proliferation and induce apoptosis of cervical cancer cells,and its regulatory mechanism may be related to the expression of Bcl-2 family proteins and the inhibition of p53 signaling pathway.
5.Surgical treatment of recurrent pseudocyst of auricle.
Jia LI ; Email: 15119331555@126.COM. ; Duoxian LI ; Xiongwen LI ; Pengjian ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2015;50(10):851-853
OBJECTIVETo investigate the surgical techniques and skills for recurrent pseudocyst of auricle.
METHODSRetrospective analysis of 20 cases from January 2006 to May 2014 in our department, who were treated by the operations with skills as follows, resection the anterior wall of pseudocyst cartilage and perichondrium, grooves cutting the roughing the posterior cartilages wall, and local compression.
RESULTSAll 20 patients were followed up for 8 to 42 months and all were cured. No recurrence, infections and malformation were happened.
CONCLUSIONResection the anterior cartilage wall perichondrium of the pseudocyst, grooves cutting and roughing of posterior cartilages of the peudocyst, and in combined with local compression are effective treatment for recurrent cases.
Cysts ; surgery ; Ear Auricle ; pathology ; Ear Diseases ; surgery ; Humans ; Recurrence ; Retrospective Studies ; Treatment Outcome
6. Effect of skin soft tissue expansion on repair of large area of scars on extremities
Junqiang PENG ; Yangqun LI ; Muxin ZHAO ; Zhe YANG ; Wen CHEN ; Yong TANG ; Ning MA ; Lisi XU ; Jiangting CHEN
Chinese Journal of Burns 2019;35(4):308-310
Objective:
To investigate the effect of skin soft tissue expansion on repair of large area of scars on extremities.
Methods:
Twenty-five patients with large area of scars on extremities were admitted to our department from June 2007 to October 2014. There were 14 males and 11 females, aged 4 to 36 years. Operations were performed under local infiltration anesthesia or general anesthesia. In the first stage, 1 to 5 cylindrical expanders with capacities of 250 to 600 mL were placed at left or right sides or at upper or lower parts of the scars. In the second stage, scars of 21 patients were repaired with expanded transverse propulsive and lateral flaps, and scars of 4 patients were repaired with expanded perforator flaps whose pedicles were perforators of brachial artery, superior ulnar collateral artery, or posterior interosseous artery according to areas and shapes of the scars. The secondary wound areas ranged from 13 cm×7 cm to 34 cm×18 cm after dissolution or excision of scars. The areas of flaps ranged from 13 cm×7 cm to 20 cm×12 cm. The donor sites were sutured directly. The flaps after operation and follow-up of patients were observed and recorded.
Results:
All expanded flaps survived after operation. And the superficial distal part of flap whose pedicle was perforator of posterior interosseous artery in one patient was with necrosis, and other flaps survived well. During follow-up of 3 to 15 months after operation of the second stage, color and texture of flaps were similar to surrounding skin, while extremities of donor sites were thinner and auxiliary incisional scars formed after expansion.
Conclusions
Expanded flap is a good way to repair large area of scar on extremities. Bilateral skin of scar is the first choice of donor site of expanded flap. If there isn′t enough skin for expanding on bilateral sides, expanded perforator flap designed at upper or lower part of the scar is another choice to repair the scar.
7.Serological Characteristics of Patients with Autoimmune Hemoly-tic Anemia and Efficacy and Safety of Incompatible Transfusion of Red Blood Cells.
Li-Na ZHANG ; Xiang-Fu LIU ; Yuan LI
Journal of Experimental Hematology 2019;27(3):916-919
OBJECTIVE:
To explore the serological characteristics of patients with autoimmune hemolytic anemia(AIHA) and analyze its clinical efficacy and safety of incompatible red blood cell transfusion.
METHODS:
Sixty AIHA patients admitted in our hospital from January 2014 to January 2018 were selected. The blood type serological characteristics of 60 patients were analyzed retrospectively. According to the type of autoantibody and the composition of different red blood cells, the efficacy and safety of erythrocyte infusions were evaluated respectively.
RESULTS:
The screen results of irregular antibody in 60 AIHA patients were positive, and the direct anti-human globulin test also was positive, including 8 cases of cold autoantibodies (13.33%), 49 cases of IgG warm autoantibodies (81.67%), and 3 cases of warm cold double autoantibodies (5%). The irregular anti-body identification test confirmed the existence of homoantiboby in 17 cases (28.33%). Out of 60 cases 34 received incompatible red blood cell (RBC) infusion for 108 time including ABO homotype non washing RBC (81 tirnes) and O type washing RBC (27 times). The infusion results showed that the total [JP2]effective rate was 57.41(62/108), total partial effective rate was 14.81% (16/108) and total ineffective rate was 27.78% (30/108).The infusion of ABO homotype non-washing RBC for 81 time showed that the effective rate was 58.02%[JP] (47/81) , partial effective rate was 12.35 (10/81) and ineffective rate was 29.67% (24/81); the infusion of O type washing RBC for 27 times showed that the effective rate was 55.56% (15/27), partial effective rate was 22.22% (6/27) and ineffective rate was 22.22% (6/27), there was no significant difference in effective rate between 2 kinds of infusion (P>0.05). The comparison of different antibody type infusion showed that in the infusion of IgM cold autoantiboay for 12 times, the effective rate was 41.67% (5/12), partial effective rate was 33.33% (4/12) and ineffective rate was 25% (3/12); in the infusion of IgG warm antoantibody for 93 times. The effective rate was 58.06% (54/93),partial effective rate was 12.90% (12/93) and ineffective rale was 29.04% (27/93), there was also no significant difference in effective rate between 2 kinds of infusion(P>0.05). However, in infusion of cold/warm double autoantibody for 3 times, the effective rate was 100% (3/3), moreover, the hemotytic reaction of infusion was not observed during the treatment .
CONCLUSION
The infusion of ABO homotype non-washing RBC and O type washing RBC both possess the high safely and efficacy for treatment of patients with AIHA, but the use of ABO homotype non-washing RBC can effectively avoid the excessive use of O type washing RBC.
Anemia, Hemolytic, Autoimmune
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Autoantibodies
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Erythrocyte Count
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Erythrocytes
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Humans
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Retrospective Studies
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Tics
8.Membrane peeling combined with intravitreal injection of bevacizumab for treatment of macular epiretinal membrane: analysis of 33 cases.
Zhiqiang LI ; Guoming ZHANG ; Kangjin SU ; Xiangmei SONG ; Ruyin TIAN ; Xunqing GU
Journal of Southern Medical University 2014;34(8):1207-1209
OBJECTIVETo assess efficacy of membrane peeling combined with intravitreal injection of bevacizumab in the treatment of macular epiretinal membrane.
METHODSFrom January, 2012 to June, 2013, 33 patients (33 eyes) with the diagnosis of macular epiretinal membrane underwent vitreous surgery and membrane peeling. The patients were randomly divided into intravitreal bevacizumab group (IVB group) and non-intravitreal bevacizumab group (non-IVB group). All the patients underwent standard three-port vitrectomy and peeling of epiretinal membrane, with intravitreal injection of 1.5 mg bevacizumab at the end of operation in IVB group. The best corrected visual acuity and optical coherence tomography (OCT) were examined before and after the treatment. The patients were followed up for 3-14 months (mean 6.5 months).
RESULTSMacular epiretinal membranes were successfully peeled during operation in all the patients without postoperative intraocular infection or bleeding. Fifteen eyes received vitrectomy combined with intravitreal injection of bevacizumab, and 18 underwent only vitreous operation and membrane peeling. At the end of the follow up, the visual acuity improved in 11 eyes (73.3%) in IVB group, as compared to 13 eyes (72.2%) in the non-IVB group (P=0.627). Central macular thickness decreased by 143∓62 µm in IVB group and by 96∓28 µm in non-IVB group, showing a significant difference between the two groups (t=5.564, P<0.01).
CONCLUSIONVitrectomy and membrane peeling combined with intravitreal injection of bevacizumab can promote the recovery of macular morphology but not visual function, and its clinical use still needs to be tested in a long-term and large-sample randomized controlled study.
Angiogenesis Inhibitors ; administration & dosage ; therapeutic use ; Antibodies, Monoclonal, Humanized ; administration & dosage ; therapeutic use ; Bevacizumab ; Epiretinal Membrane ; drug therapy ; pathology ; Humans ; Intravitreal Injections ; Postoperative Complications ; Tomography, Optical Coherence ; Treatment Outcome ; Visual Acuity ; Vitrectomy ; Vitreous Body ; surgery
9.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
10.Study on the chemical composition differences of different parts of Alpinia oxyphylla Miq.based on multivariate statistical analysis
Wuyi LI ; Meizhou LI ; Ting CUI ; Yanzhen LI ; Yao HUANG ; Sen HUANG ; Zhipeng ZHANG
International Journal of Traditional Chinese Medicine 2022;44(7):776-780
Objective:To establish the Ultra-High Performance Liquid Chromatography (UPLC) characteristic chromatogram of different parts of Alpinia oxyphylla Miq., and to compare different parts of the chemical components based on multivariate statistical analysis. Methods:The UPLC was used to establish the fingerprint of Alpinia oxyphylla Miq. . The chromatograms were matched to generate the UPLC charactersistic chromatogram of different parts. Based on the variance analysis of single factor, combined with the Principal Component Analysis (PCA) ,Cluster Analysis (CA) and the Partial Orthogonal Least Square Discriminant Analysis (OPLS-DA) to analyze the differences of different medicinal parts of Alpinia oxyphylla Miq.. Results:16 common peaks of Alpinia oxyphylla Miq. were demarcated in crude drugs, compared with the medicinal materials of Alpinia oxyphylla Miq., the peak 13 (tectochhrysin) was lost in the decoction pieces, and the shell were missing peak 5 and peak 6. The results of PCA and CA showed that 15 batches of different medicinal parts of Alpinia oxyphylla Miq. can be broadly divided into 3 categories. The OPLS-DA result showed that the value of the peak area of peaks 14 (Nootkatone), 4, 7 and 12 were the main factors affecting the chemical composition of different parts of Alpinia oxyphylla Miq. .Conclusion:The fingerprint determination method established in this study is stable and controllable, which could distinguish the different parts of Alpinia oxyphylla Miq. .