1.Identification of endothelial cell key genes associated with pathogenesis and invasion of human venous malformations using single-nucleus RNA sequencing-based co-expression network analysis
Wenbo LIU ; Junjie LIN ; Meijuan ZHANG ; Chunjie YUAN ; Xiaojuan FENG ; Wenting JIAO ; Junbo QIAO ; Wenqiu WANG ; Bin FANG ; Changkuan CHEN
Chinese Journal of Preventive Medicine 2025;59(4):458-467
Objective:This study aimed to identify key genes in endothelial cell (EC) associated with the pathogenesis and progression of human venous malformations (VMs) through bioinformatics analysis, providing potential biomarkers for early screening and targeted therapy of VMs.Methods:A case-control study was conducted using surgically resected tissue specimens from VMs patients at the Third Affiliated Hospital of Zhengzhou University (from September 2021 to September 2023), with malformed venous tissues as the experimental group and distal normal venous tissues as controls. Single-nucleus RNA sequencing (snRNA-seq) was performed on paired experimental and control samples from four VM patients. High-dimensional weighted gene co-expression network analysis (hdWGCNA), combined with gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein-protein interaction (PPI) network analysis, identified critical genes. Validation experiments included 15 additional VM cases and controls using reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and Western blot.Results:A total of 55 430 nuclei were captured using snRNA-seq, with 30 391 nuclei from the experimental group and 25 039 nuclei from the control group. Cluster analysis identified 22 distinct cell populations, which were annotated into 8 cell types. hdWGCNA revealed four modules associated with invasion, which were enriched in angiogenesis, integrin signaling, and cell adhesion according to GO analysis. KEGG pathway analysis indicated that the PI3K-AKT signaling pathway and focal adhesion are key regulatory mechanisms. PPI network analysis combined with cytoscape identified EGFL7, TEK, and FLT1 as key genes. RT-qPCR results demonstrated that the relative mRNA expression levels of these three genes in the experimental group (6.66±2.31, 1.86±0.62, 3.49±0.58) were significantly higher than those in the control group (1.05±0.14, 1.00±0.14, 1.06±0.25), with statistically significant differences ( t=9.37, 4.27, 11.20, P<0.05). Immunohistochemical analysis showed that the relative protein expression levels of these three genes in the cytoplasm of the experimental group (0.84±0.15, 0.68±0.14, 0.85±0.12) were also significantly higher than those in the control group (0.19±0.05, 0.23±0.06, 0.30±0.05), with statistically significant differences ( t=16.62, 5.93, 11.68, P<0.05). Western blot analysis confirmed that the relative protein expression levels of these three genes in the experimental group (0.35±0.04, 0.36±0.09, 0.31±0.04) were significantly higher than those in the control group (0.19±0.01, 0.13±0.02, 0.14±0.04), with statistically significant differences ( t=7.05, 4.61, 5.93, P<0.05). Conclusion:EGFL7, FLT1, and TEK in EC may play crucial roles in the occurrence and invasion of VMs.
2.Identification of endothelial cell key genes associated with pathogenesis and invasion of human venous malformations using single-nucleus RNA sequencing-based co-expression network analysis
Wenbo LIU ; Junjie LIN ; Meijuan ZHANG ; Chunjie YUAN ; Xiaojuan FENG ; Wenting JIAO ; Junbo QIAO ; Wenqiu WANG ; Bin FANG ; Changkuan CHEN
Chinese Journal of Preventive Medicine 2025;59(4):458-467
Objective:This study aimed to identify key genes in endothelial cell (EC) associated with the pathogenesis and progression of human venous malformations (VMs) through bioinformatics analysis, providing potential biomarkers for early screening and targeted therapy of VMs.Methods:A case-control study was conducted using surgically resected tissue specimens from VMs patients at the Third Affiliated Hospital of Zhengzhou University (from September 2021 to September 2023), with malformed venous tissues as the experimental group and distal normal venous tissues as controls. Single-nucleus RNA sequencing (snRNA-seq) was performed on paired experimental and control samples from four VM patients. High-dimensional weighted gene co-expression network analysis (hdWGCNA), combined with gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein-protein interaction (PPI) network analysis, identified critical genes. Validation experiments included 15 additional VM cases and controls using reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and Western blot.Results:A total of 55 430 nuclei were captured using snRNA-seq, with 30 391 nuclei from the experimental group and 25 039 nuclei from the control group. Cluster analysis identified 22 distinct cell populations, which were annotated into 8 cell types. hdWGCNA revealed four modules associated with invasion, which were enriched in angiogenesis, integrin signaling, and cell adhesion according to GO analysis. KEGG pathway analysis indicated that the PI3K-AKT signaling pathway and focal adhesion are key regulatory mechanisms. PPI network analysis combined with cytoscape identified EGFL7, TEK, and FLT1 as key genes. RT-qPCR results demonstrated that the relative mRNA expression levels of these three genes in the experimental group (6.66±2.31, 1.86±0.62, 3.49±0.58) were significantly higher than those in the control group (1.05±0.14, 1.00±0.14, 1.06±0.25), with statistically significant differences ( t=9.37, 4.27, 11.20, P<0.05). Immunohistochemical analysis showed that the relative protein expression levels of these three genes in the cytoplasm of the experimental group (0.84±0.15, 0.68±0.14, 0.85±0.12) were also significantly higher than those in the control group (0.19±0.05, 0.23±0.06, 0.30±0.05), with statistically significant differences ( t=16.62, 5.93, 11.68, P<0.05). Western blot analysis confirmed that the relative protein expression levels of these three genes in the experimental group (0.35±0.04, 0.36±0.09, 0.31±0.04) were significantly higher than those in the control group (0.19±0.01, 0.13±0.02, 0.14±0.04), with statistically significant differences ( t=7.05, 4.61, 5.93, P<0.05). Conclusion:EGFL7, FLT1, and TEK in EC may play crucial roles in the occurrence and invasion of VMs.
3.Expert consensus on the evaluation and rehabilitation management of shoulder syndrome after neek dissection for oral and maxillofacial malignancies
Jiacun LI ; Moyi SUN ; Jiaojie REN ; Wei GUO ; Longjiang LI ; Zhangui TANG ; Guoxin REN ; Zhijun SUN ; Jian MENG ; Wei SHANG ; Shaoyan LIU ; Jie ZHANG ; Jicheng LI ; Yue HE ; Chunjie LI ; Kai YANG ; Zhongcheng GONG ; Qing XI ; Bing HAN ; Huaming MAI ; Yanping CHEN ; Jie ZHANG ; Yadong WU ; Chao LI ; Changming AN ; Chuanzheng SUN ; Hua YUAN ; Fan YANG ; Haiguang YUAN ; Dandong WU ; Shuai FAN ; Fei LI ; Chao XU ; Wei WEI
Journal of Practical Stomatology 2024;40(5):597-607
Neck dissection(ND)is one of the main treatment methods for oral and maxillofacial malignancies.Although ND type is in con-stant improvement,but intraoperative peal-pull-push injury of the accessory nerve,muscle,muscle membrane,fascia and ligament induced shoulder syndrome(SS)is still a common postoperative complication,combined with the influence of radiochemotherapy,not only can cause pain,stiffness,numbness,limited dysfunction of shoulder neck and arm,but also may have serious impact on patient's life quality and phys-ical and mental health.At present,there is still a lack of a systematic evaluation and rehabilitation management program for postoperative SS of oral and maxillofacial malignant tumors.Based on the previous clinical practice and the current available evidence,refer to the relevant lit-erature at home and abroad,the experts in the field of maxillofacial tumor surgery and rehabilitation were invited to discuss,modify and reach a consenusus on the etiology,assessment diagnosis,differential diagnosis,rehabilitation strategy and prevention of SS,in order to provide clinical reference.
4.Zanthoxylum bungeanum Aqueous Extract Treated Rheumatoid Arthritis by Inhibiting Synovial Cell Proliferation and Inducing Cell Apoptosis
Ziqi LIU ; Yurou WU ; Lilin ZHANG ; Xue SUN ; Xing YUAN ; Chunjie WU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(20):30-38
ObjectiveTo investigate the therapeutic effect of the water extract of Zanthoxylum bungeanum aqueous extract(ZBAE)on rheumatoid arthritis. MethodThe sixty SD rats were divided into normal group, model group [complete Freund's adjuvant (CFA), 10 mg·kg-1], methotrexate(MTX) group (0.25 mg·kg-1), low -, medium -, and high-dose ZBAE groups (90, 180, 360 mg·kg-1). The rats in MTX group were given intraperitoneal injection for two weeks, three times a week, and the rats in ZBAE group were administrated for 14 days. The swelling of the ankle joint and body weight were observed, and arthritis scores were also performed. Computed tomography (CT), hematoxylin-eosin (HE) staining and safranine-O and fast green staining were used to observe the effect of ZBAE on synovial hyperplasia and bone protection. Cell counting kit-8(CCK-8)method was used to detect the proliferation of the RA-FLSs cells treated with ZBAE. According to the results of CCK-8 experiment, the optimal concentration and time of administration were determined, blank group, low -, medium -, and high-dose ZBAE groups (0.08,0.10,0.12 g·L-1) were set up. The cell cycle distribution and apoptosis rate were analyzed by flow cytometry,the migration ability of RA-FLSs cells was examined by scratch test. Western blot was used to detect the effect of ZBAE on phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), cyclin-dependent kinase 2 (CDK2), Cyclin A and phosphorylated PI3K, Akt (p-PI3K,p-Akt) protein expression in RA-FLSs cells. ResultCompared with the normal group,joint swelling index and arthritis score were increased in the model group (P<0.05),the bone of the ankle was seriously damaged, and there was obvious synovial hyperplasia. Compared with the model group, the ZBAE group could significantly reduce the joint swelling index (P<0.05), inhibit synovial hyperplasia and cartilage destruction. In vitro study showed that compared with the blank group, ZBAE could inhibit the migration of RA-FLSs (P<0.05), promoted cell apoptosis (P<0.05), and acted on RA-FLSs cells in S phase to inhibit cell proliferation. Moreover, the result of Western blot showed that compared with the blank group, the expression of p-PI3K, p-Akt, CDK2 and Cyclin A proteins were significantly decreased in the high dose group of ZBAE (P<0.05). ConclusionThese results suggest that ZBAE has a therapeutic effect on rheumatoid arthritis by inhibiting synovial hyperplasia, promoting synovial apoptosis and inhibiting its migration.
5.Ultrasonographic characteristics of the follicular variant of papillary thyroid cancer in children and adolescents
Jiangyan LOU ; Junping LIU ; Yuan CHEN ; Haimiao XU ; Zhenying GUO ; Chunjie HOU ; Dong XU ; Lingyan ZHOU ; Liyu CHEN
Chinese Journal of Endocrine Surgery 2019;13(2):135-138
Objective To compare the sonographic features as well as clinical histopathological features of follicular variant papillary thyroid carcinoma(FVPTC) and conventional papillary thyroid carcinoma (CPTC) in pediatric patients.Methods From Jan.2006 to Dec.2017,26 FVPTC patients and 82 CPTC patients were enrolled in this study.The clinical histopathological findings and the sonographic features were compared between the two groups.FVPTCs and CPTCs were divided into PTC-like and follicular neoplasm(FN)-like based on sonographic characteristics.Results The mean nodule size of FVPTCs was larger than that of conventional PTCs.Extrathyroid invasion and cervical lymph node metastasis did not have significant difference between CPTC and FVPTC patients(53.8% vs 62.2% and 76.9% vs 82.9%,respectively).Multiple nodules(P=0.000)and distant pulmonary metastases(P=0.024) were more frequent in CPTCs than in FVPTCs(P<0.05).The rate of an ill-defined margin (P=0.000) and calcification (P=0.003)in terms of sonographic features were lower in FVPTCs than conventional PTCs(P<0.05).A Ⅴ+Ⅵ diagnosis of PTC on FNAC of FVPTCs was less common than that of conventional PTCs (P=0.014).Multifocality(P=0.000),extrathyroidal invasion (P=0.000),and lymph node metastasis (P=0.000) were significantly different between PTC-like FVPTCs and FN-like FVPTCs.Conclusion FVPTC in children and adolescents shows a relatively larger size,more benign sonographic features,and a lower diagnostic rate of PTC by FNAC compared with conventional PTCs in pediatric patients.
6.Construction of Lactobacillus rhamnosus GG particles surface display system.
Runyu SU ; Boyao NIE ; Shengling YUAN ; Haoxia TAO ; Chunjie LIU ; Bailiang YANG ; Yanchun WANG
Chinese Journal of Biotechnology 2017;33(1):132-140
To describe a novel particles surface display system which is consisted of gram-positive enhancer matrix (GEM) particles and anchor proteins for bacteria-like particles vaccines, we treated Lactobacillus rhamnosus GG bacteria with 10% heated-TCA for preparing GEM particles, and then identified the harvested GEM particles by electron microscopy, RT-PCR and SDS-PAGE. Meanwhile, Escherichia coli was induced to express hybrid proteins PA3-EGFP and P60-EGFP, and GEM particles were incubated with them. Then binding of anchor proteins were determined by Western blotting, transmission electron microscopy, fluorescence microscopy and spectrofluorometry. GEM particles preserved original size and shape, and proteins and DNA contents of GEM particles were released substantially. The two anchor proteins both had efficiently immobilized on the surface of GEM. GEM particles that were bounded by anchor proteins were brushy. The fluorescence of GEM particles anchoring PA3 was slightly brighter than P60, but the difference was not significant (P>0.05). GEM particles prepared from L. rhamnosus GG have a good binding efficiency with anchor proteins PA3-EGFP and P60-EGFP. Therefore, this novel foreign protein surface display system could be used for bacteria-like particle vaccines.
7."Current Situation and Thinking on ""Odor and Taste"" Identification of Traditional Chinese Medicine"
Shilong YANG ; Na WU ; Xing YUAN ; Yujie LIU ; Ruixue ZHONG ; Chunjie WU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(9):1876-1879
Odor and taste are characteristic properties of traditional Chinese medicine (TCM). They are closely related to the chemical compositions in TCM. They are also the outer reflection of TCM quality and an important part of the character identification. This article reviewed the current research on odor and taste identification of TCM, which indicated that odor and taste identification had not been paid enough attention to and its research lagged behind modern social development. Based on current analytical techniques and methods, this article proposed a new idea for odor and taste identification of TCM. The idea emphasized the objectification of odor and taste and TCM identification research should be combined with chemical components analysis.
8.Systematic review on control of swelling and trismus after extraction of impacted mandibular third molar by dexamethasone pericoronal injection.
Chunjie LI ; Hongwei ZHAO ; Longjiang LI ; Shuangjun LI ; Yuan WU ; Xuejuan LIAO ; Jian PAN
West China Journal of Stomatology 2013;31(3):267-271
OBJECTIVETo assess the efficacy of Dexamethasone (DM) pericoronal injection for the control of swelling and trismus caused by impacted mandibular third molars extraction.
METHODSCochrane, PUBMED, EMBASE and CBM were searched for eligible studies. Hand-searching included references of the included studies and Chinese dental journals. Risk of bias of the included studies was assessed by two reviewers independently using Cochrane Collaboration's tool, and data extraction was done by them. Meta-analysis was delivered with Revman 5.1.
RESULTSSeven randomized controlled trials, involving 684 participants, were included. Six of them had moderate risk of bias and one had high risk of bias. Meta-analysis showed that DM pericoronal injection could relieve trismus by 6.77 mm (P=0.02) within 1-2 days after the surgery. It could also reduce 51% of the risk of moderate-severe trismus(P<0.000 01) and could significantly control facial swelling (P<0.05). There was no differences between 4 mg and 8 mg DM (P>0.05).
CONCLUSIONPeriodontal injection of 4-5 mg DM could control facial swelling and trismus following impacted mandibular third molar extraction. But more randomized controlled trials are needed.
Dexamethasone ; Edema ; Humans ; Mandible ; Molar, Third ; Pain, Postoperative ; Tooth Extraction ; Tooth, Impacted ; Trismus
9.Drainage for the control of complications after extraction of impacted mandibular third molar: a systematic review.
Shuangjun LI ; Chunjie LI ; Yuan WU ; Jian PAN ; Hongwei ZHAO ; Longjiang LI
West China Journal of Stomatology 2012;30(6):615-619
OBJECTIVETo assess the efficacy and safety of drainage for the control of the complications following impacted mandibular third molar extraction.
METHODSTo retrieve randomized controlled trials assessing the efficacy and safety of drainage for the control of the complications following impacted mandibular third molar extraction, bibliographic databases including MEDLINE, Cochrane Controlled Trials Register, EMBASE, OPEN SIGLE and China Biology Medicine Database (CBM) were searched on August 23th 2011. References of the included studies and Chinese dental journals were hand-searched. The risk of bias were used by Cochrane Collaboration's tool and the data were extracted. Meta-analysis was done with Revman 5.1.
RESULTSNine articles met the eligibility criteria and were included, seven randomized controlled trials and 2 quasi-randomized controlled trials. Seven of these studies had unclear risk of bias and 2 had high risk of bias. Drainage could significantly increase 4.44 mm of the post-operative maximal mouth opening (P = 0.003), relief facial swelling (P < 0.05) and reduce post-operative complications (P = 0.008). But no evidence showed that drainage had a positive effects on post-operative pain (P = 0.09).
CONCLUSIONDrainage could probably control the complications following impacted mandibular third molar extraction; but more randomized controlled trials are needed to reinforce the conclusion.
China ; Drainage ; Humans ; Mandible ; Molar, Third ; Postoperative Complications ; Tooth Extraction
10.Development of a killed but metabolically active anthracis vaccine candidate strain.
Fei SHEN ; Shengling YUAN ; Dewen ZHAN ; Yanchun WANG ; Min REN ; Haoxia TAO ; Peng WANG ; Lingchun WANG ; Dongsheng CHEN ; Chunjie LIU
Chinese Journal of Biotechnology 2011;27(5):781-789
Anthrax is a zoonosis caused by Bacillus anthracis, which seriously affects human health. In recent years, a special phenomenon is found that the metabolic active of a bacterium remains after it is killed. To development of a KBMA (killed but metabolically active) Bacillus anthracis vaccine candidate strain, a plasmid pMAD and a recombinase system Cre-loxP were used to knockout the uvrAB gene of B. anthracis AP422 which lacks both of two plasmids pXO1 and pXO2. The results of PCR and RT-PCR shows that uvrAB genes were deleted from B. anthracis AP422 chromosome successfully. The constructed B. anthracis AP422deltauvrAB was inactivated by photochemical treatment (PCT) including an exposure in a long-wave-length ultraviolet (UVA) light and a treatment of 8-Methoxypsoralen (8-MOP), then the metabolic activity were detected by the method of MTS. The results showed that the killed B. anthracis AP422deltauvrAB maintained a highly metabolic activity for at least 4 hours, showing a state of KBMA. The KBMA strain of B. anthracis AP422deltauvrAB provides the prospective vaccine candidate strain for anthrax.
Anthrax
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immunology
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microbiology
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prevention & control
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Anthrax Vaccines
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genetics
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immunology
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radiation effects
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Bacillus anthracis
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genetics
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immunology
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Gene Knockout Techniques
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Methoxsalen
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pharmacology
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Ultraviolet Rays
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Vaccines, Inactivated
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genetics
;
immunology

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