1.Effect of high-dose vitamin B6 on stress-induced liver cell death in rats with severe trauma and the possible mechanism
Yinjie ZHANG ; Zhihuai WANG ; Xuelin TANG ; Haiyang ZHOU ; Peng GAO ; Chunfu ZHU ; Zhongzhi JIA ; Maoxing YUE ; Xihu QIN
Chinese Journal of Trauma 2024;40(6):558-568
Objective:To investigate the effect of high-dose vitamin B6 on stress-induced liver cell death in rats with severe trauma and its possible mechanism.Methods:Thirty-two male SD rats were selected and divided into sham surgery group, sham surgery+B6 group, trauma group, and trauma+B6 group by using a random number table, with 8 rats in each group. Rat models of severe trauma were established by inducing abdominal wall injury, bilateral femoral fractures, unilateral cranial injury, and withdrawal of 4 ml blood from the femoral artery. The sham surgery+B6 group and trauma+B6 group were treated with saline solution plus high-dose vitamin B6, while the sham surgery group and trauma group with infusion of saline solution only. At 36 hours after injury, rat liver tissues were collected for the following experiments: (1) the genes differentially expressed in the liver tissues of the rats of the trauma group and the trauma+B6 group were screened with next-generation sequencing, followed by an analysis of the possible involvement of cell death pathways; (2) validation was conducted to ascertain whether high-dose vitamin B6 could influence various cell death pathways in the liver cells in the sham surgery group, sham surgery+B6 group, trauma group, and trauma+B6 group: apoptosis was confirmed through terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining; necroptosis was verified by mixed lineage kinase domain-like protein (MLKL) immunohistochemical staining; autophagy was examined via transmission electron microscopy; ferroptosis was confirmed by detecting oxidative malondialdehyde (MDA) levels, oxidized glutathione levels, Prussian blue staining with diaminobenzidine (DAB) enhancement, transmission electron microscopy, and immunohistochemical staining for acyl-CoA synthetase long-chain family member 4 (ACSL4); (3) Biological information analyses [Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Enrichment analysis (GSEA)] were performed for biological processes and signaling pathways represented by liver tissue sequencing results of rats between the trauma group and the trauma+B6 group.Results:(1) In the liver tissues of rats, there were 344 significantly differentially expressed genes between the trauma group and trauma+B6 group, comprising 137 upregulated genes and 207 downregulated genes, of which 18 genes were associated with apoptosis, autophagy, necroptosis, ferroptosis, and pyroptosis. (2) No significant differences were found in TUNEL staining among the sham surgery group, sham surgery+B6 group, trauma group or trauma+B6 group; MLKL protein expression levels in the liver tissues after trauma were improved, of which the trauma+B6 group was lower than that of the trauma group; Electron microscopy showed that autophagic activity in the liver cells were significantly increased after trauma, which was significantly lower of the trauma+B6 group than that of the trauma group; MDA levels in the rat liver tissues were (0.20±0.05)nmol/mg, (0.17±0.07)nmol/mg, (0.69±0.11)nmol/mg and (0.52±0.07)nmol/mg in the sham surgery group, sham surgery+B6 group, trauma group, and trauma+B6 group respectively ( P<0.01), with the trauma group having the highest MDA levels and trauma+B6 group having lower MDA levels than the trauma group; Oxidized glutathione levels in the liver tissues of the four groups were (11.75±2.09)μmol/g, (11.69±1.66)μmol/g, (19.75±3.40)μmol/g, and (14.51±1.46)μmol/g respectively ( P<0.01), with the trauma group having the highest levels and trauma+B6 group having lower levels than the trauma group; Significantly increased iron deposition was observed in the liver tissues after trauma, with lower iron deposition in trauma+B6 group than the trauma group; Electron microscopy revealed significantly lower mitochondrial membrane density in the trauma+B6 group compared to the trauma group. ACSL4 protein expression level was lower in the trauma+B6 group compared to the trauma group; (3) GO, KEGG and GSEA enrichment analyses suggested that high-dose vitamin B6 may enhance cholesterol synthesis metabolism in the liver cells and alleviate oxidative stress to reduce liver cell damage and restore normal liver cell function after trauma. Conclusions:High-dose vitamin B6 attenuates stress-induced liver injury in rats with severe trauma by inhibiting the progression of necroptosis, autophagy and ferroptosis. Its molecular mechanism may be associated with enhanced hepatic cholesterol synthesis metabolism and alleviation of oxidative stress in the liver cells.
2.Double heterozygous pathogenic mutations in KIF3C and ZNF513 cause hereditary gingival fibromatosis.
Jianfan CHEN ; Xueqing XU ; Song CHEN ; Ting LU ; Yingchun ZHENG ; Zhongzhi GAN ; Zongrui SHEN ; Shunfei MA ; Duocai WANG ; Leyi SU ; Fei HE ; Xuan SHANG ; Huiyong XU ; Dong CHEN ; Leitao ZHANG ; Fu XIONG
International Journal of Oral Science 2023;15(1):46-46
Hereditary gingival fibromatosis (HGF) is a rare inherited condition with fibromatoid hyperplasia of the gingival tissue that exhibits great genetic heterogeneity. Five distinct loci related to non-syndromic HGF have been identified; however, only two disease-causing genes, SOS1 and REST, inducing HGF have been identified at two loci, GINGF1 and GINGF5, respectively. Here, based on a family pedigree with 26 members, including nine patients with HGF, we identified double heterozygous pathogenic mutations in the ZNF513 (c.C748T, p.R250W) and KIF3C (c.G1229A, p.R410H) genes within the GINGF3 locus related to HGF. Functional studies demonstrated that the ZNF513 p.R250W and KIF3C p.R410H variants significantly increased the expression of ZNF513 and KIF3C in vitro and in vivo. ZNF513, a transcription factor, binds to KIF3C exon 1 and participates in the positive regulation of KIF3C expression in gingival fibroblasts. Furthermore, a knock-in mouse model confirmed that heterozygous or homozygous mutations within Zfp513 (p.R250W) or Kif3c (p.R412H) alone do not led to clear phenotypes with gingival fibromatosis, whereas the double mutations led to gingival hyperplasia phenotypes. In addition, we found that ZNF513 binds to the SOS1 promoter and plays an important positive role in regulating the expression of SOS1. Moreover, the KIF3C p.R410H mutation could activate the PI3K and KCNQ1 potassium channels. ZNF513 combined with KIF3C regulates gingival fibroblast proliferation, migration, and fibrosis response via the PI3K/AKT/mTOR and Ras/Raf/MEK/ERK pathways. In summary, these results demonstrate ZNF513 + KIF3C as an important genetic combination in HGF manifestation and suggest that ZNF513 mutation may be a major risk factor for HGF.
Animals
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Humans
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Mice
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Fibromatosis, Gingival/pathology*
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Gingiva
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Kinesins/genetics*
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Mutation/genetics*
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Phosphatidylinositol 3-Kinases/genetics*
3.Quality Evaluation of Physical Properties of Ultrafine Granular Powder of Salviae Miltiorrhizae Radix et Rhizoma
Qian-liang ZHANG ; Zheng LUO ; Wen DENG ; Jin-le CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(23):145-152
Objective:Taking ultrafine granular powder of Salviae Miltiorrhizae Radix et Rhizoma (UGPSMR) as the research object, to establish a method for evaluating its physical properties. Method:A method was established for measuring the particle size distribution and specific surface area of UGPSMR, and the methodological investigation was carried out. A total of 15 physical indicators [
4.Quality evaluation of angelica broken wall powder based on QbD concept of traditional Chinese medicine.
Zheng LUO ; Wen DENG ; Qian-Liang ZHANG ; Bi-Yan FANG ; Jin-le CHENG
China Journal of Chinese Materia Medica 2020;45(5):1105-1113
In this paper, angelica broken wall powder(ABWP) was taken as the research object, HPLC fingerprint combined with multi-component determination(ferulic acid, senkyunolide I, coniferyl ferulate, ligustilide and 3-butylidenephthalide), physical fingerprint(D_(90), particle size distribution range, particle size distribution width, bulk density, tap density, inter-particle porosity, Carr index, specific surface area, pore volume, angle of repose, Hausner ratio, loss on drying and hygroscopicity)were used to characterize the quality attribute of ABWP; similarity analysis, cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were conducted to construct the quality evaluation method of holographic analysis based on traditional Chinese medicine QbD "4 H mode", in order to evaluate the quality of ABWP from different sources and find out differentiated indicators. The quality evaluation method could be used for scientific, comprehensive evaluation of the quality attribute of ABWP, and the quality consistency evaluation of cell-wall-broken powder of different sources or different processes.It provides new ideas for quality control and research of ultrafine granular powders of traditional Chinese medicine.
Angelica/chemistry*
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal/analysis*
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Medicine, Chinese Traditional
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Powders
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Quality Control
5.DPHL:A DIA Pan-human Protein Mass Spectrometry Library for Robust Biomarker Discovery
Zhu TIANSHENG ; Zhu YI ; Xuan YUE ; Gao HUANHUAN ; Cai XUE ; Piersma R. SANDER ; Pham V. THANG ; Schelfhorst TIM ; Haas R.G.D. RICHARD ; Bijnsdorp V. IRENE ; Sun RUI ; Yue LIANG ; Ruan GUAN ; Zhang QIUSHI ; Hu MO ; Zhou YUE ; Winan J. Van Houdt ; Tessa Y.S. Le Large ; Cloos JACQUELINE ; Wojtuszkiewicz ANNA ; Koppers-Lalic DANIJELA ; B(o)ttger FRANZISKA ; Scheepbouwer CHANTAL ; Brakenhoff H. RUUD ; Geert J.L.H. van Leenders ; Ijzermans N.M. JAN ; Martens W.M. JOHN ; Steenbergen D.M. RENSKE ; Grieken C. NICOLE ; Selvarajan SATHIYAMOORTHY ; Mantoo SANGEETA ; Lee S. SZE ; Yeow J.Y. SERENE ; Alkaff M.F. SYED ; Xiang NAN ; Sun YAOTING ; Yi XIAO ; Dai SHAOZHENG ; Liu WEI ; Lu TIAN ; Wu ZHICHENG ; Liang XIAO ; Wang MAN ; Shao YINGKUAN ; Zheng XI ; Xu KAILUN ; Yang QIN ; Meng YIFAN ; Lu CONG ; Zhu JIANG ; Zheng JIN'E ; Wang BO ; Lou SAI ; Dai YIBEI ; Xu CHAO ; Yu CHENHUAN ; Ying HUAZHONG ; Lim K. TONY ; Wu JIANMIN ; Gao XIAOFEI ; Luan ZHONGZHI ; Teng XIAODONG ; Wu PENG ; Huang SHI'ANG ; Tao ZHIHUA ; Iyer G. NARAYANAN ; Zhou SHUIGENG ; Shao WENGUANG ; Lam HENRY ; Ma DING ; Ji JIAFU ; Kon L. OI ; Zheng SHU ; Aebersold RUEDI ; Jimenez R. CONNIE ; Guo TIANNAN
Genomics, Proteomics & Bioinformatics 2020;18(2):104-119
To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipe-line and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to gen-erate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.
6.Research on preparation and relevant performance test for new-type acellular dermal matrix
Wu XIONG ; Biao ZHANG ; Xu CAI ; Xinling HUANG ; Qiaoli HUANG ; Wenjuan QUAN ; Yingying CHEN ; Hongwei LAN ; Zhongzhi ZHOU
Journal of Chinese Physician 2017;19(7):1018-1021
Objective To prepare a new-type acellular dermal matrix (ADM) and research on its relevant performance,which would provide theoretical evidence for clinical application.Methods Skin of Bama suckling pig was taken as resource of skin,and technologies of physics,chemistry and biology were selected to prepare new-type ADM.To detect the external structure,physical and chemical property as well as biological property of the prepared new-type ADM,hematoxylin-eosin (HE) staining observation,scanning electron microscope observation,amino acid analysis,material porosity and hydrophilicity test,tensile strength and in vitro degradation experiment,cytotoxicity test,and animal experiment have been conducted.Results New-type ADM cells have been thoroughly removed and dermal matrix remains intact with collagen content of 95.55%,connective three-dimensional pore structure,(85.03 ± O.99) % of porosity,(24.56 ± 0.57) ° of contact angle implying new-type ADM was hydrophilic substance,(5.48 ± 0.44) Pa of tensile strength implying its moderate level of pulling force,in vitro degradation period reduced to (28.7 ± O.76) h,and >75% relative growth rate (RGR).Cells grew and proliferated on new-type ADM and could be replaced by original tissue after degradation.Conclusions New-type ADM have overcome disadvantages of traditional preparation method in sabotaging dermal matrix structure and incompletely removing cells from matrix,which is qualified with higher level of collagen content and porosity.With improved biological property,greatly reduced inflammation immunoreactions,and accelerated degradation rate,new-type ADM is of higher level of clinical application value.
7.The study on correlation between PROC gene mutation and venous thrombosis
Zhigang ZHAO ; Wenkai ZHANG ; Bo LI ; Qing CHENG ; Guoguo ZHU ; Zhongzhi TANG
The Journal of Practical Medicine 2016;32(3):421-424
Objective To study the effect of protein C mutation on venous thrombosis (VTE) by PROC gene sequencing in patients with VTE. Methods Human PROC gene sequence was designed to amplify the third exon region of the PC-Gla domain , and then the PCR products were sequenced to search for a single nucleotide mutation (SNVs). The SNVs was constructed into eukaryotic expression system and a stable expression of wild-type and mutant PC cells were also constructed. At the same time , the distribution of PC levels in normal and VTE patients were detected with ELISA. Results Three single nucleotide mutations were found in different patients. In HUVEC cells, the synthesis of PC decreased in each mutant strain. The PC level in the normal patients , VTE patients , and the mutant samples were detected , which were significantly lower in the mutant samples than that of the VTE group (P = 0.035 3) and the normal level (P < 0.000 1). Conclusion Three mutation sites PCArg-1Cys , PCVal34Met and PCArg9Cys are important genetic factors lead to a significant decrease in plasma PC levels and the increase of VTE risk.
8.Catheter aspiration alone or combined with thrombolysis in the treatment of superior mesenteric artery embolism.
Yuanquan HUANG ; Zhongzhi JIA ; Qi WANG ; Wenhua CHEN ; Zhongming HE ; Jun ZHANG ; Kai WANG ; Feng TIAN
Chinese Journal of Gastrointestinal Surgery 2014;17(10):1018-1021
OBJECTIVETo investigate the efficacy of catheter aspiration or combined with thrombolysis in the treatment of superior mesenteric artery embolism(SMAE).
METHODSClinical and imaging data of 25 SMAE patients who underwent catheter aspiration or combined with urokinase thrombolysis in the First People's Hospital and the Second People's Hospital of Changzhou from January 2005 to July 2013 were retrospectively analyzed.
RESULTSTwenty patients were confirmed as SMA trunk embolism and 5 as SMA branch artery embolism. The embolic SMA trunks were completely recannulated by catheter aspiration in the above 20 cases, but small emboli embolized distal branch artery in 6 cases. These 6 patients plus above 5 patients with branch artery embolism received catheter aspiration combined with thrombolytic therapy. Among these 11 patients, complete open, partial open and non-open of branch arteries were found in 5, 3, 3 cases respectively, while collateral circulation increased significantly in non-open patients. During the follow-up period of (4.1±2.2) months, clinical symptom relief and digestive function recovery were observed in 24 cases. Only one case underwent bowel resection because of intestinal necrosis 24 hours after treatment and developed short bowel syndrome.
CONCLUSIONCatheter aspiration or combined with thrombolysis is a safe and effective method in treating SMAE.
Embolism ; therapy ; Humans ; Intestinal Diseases ; Mesenteric Artery, Superior ; Mesenteric Vascular Occlusion ; therapy ; Retrospective Studies ; Thrombolytic Therapy
9.Analysis on the results of etiology and serology of plague in Qinghai province from 2001 to 2010.
Yonghai YANG ; Mei WANG ; Xiaolong ZHAO ; Zhongzhi ZHAO ; Aiping ZHANG ; Rongjie WEI ; Baiqing WEI ; Zuyun WANG
Chinese Journal of Preventive Medicine 2014;48(2):124-127
OBJECTIVETo analyze the results of etiology and serology of plague among human and infected animals in Qinghai province from 2001 to 2010.
METHODSThirty-seven cases of human infected with plague, 53 541 different animal samples, 5 685 sets of vector insects flea and 49 039 different animal serum samples were obtained between 2001 and 2010. A total of 7 811 samples of serum from healthy farmers and herdsmen in 14 counties in Qinghai from 2005 to 2007 were collected. Yersinia pestis (Y. pestis) were detected in visceral and secretions from human, infected animals and vector insects, respectively. Plague antigen was detected by reverse indirect hemagglutination assay (RIHA) in those samples. Indirect hemagglutination assay (IHA) was used to test plague FI antibody in serum of human and infected animals.
RESULTS37 human plague cases were confirmed, 21 strains of plague Y. pestis were isolated from human cases and 14 positive were detected out. 133 of 7 811 samples of human serum were IHA positive, with the positive rate at 1.7%. A total of 146 strains of plague were isolated from infected animals and vector insects, 99 out of which were from infected animals, with a ratio of Marmota himalayan at 72.7% (72/99) and the other 47 were from vector insects, with a ratio of callopsylla solaris at 68.1% (32/47). The number of IHA and PIHA positive were 300 and 10, respectively. A total of 3 animals and 3 insects species were identified as new epidemic hosts for plague. The natural plague focus of Microtus fuscus was discovered and confirmed and coexisted with natural focus of Marmota himalayan in Chengduo county, Yushu prefecture. The epidemic situation of plague is distributed mainly in Haixi, Yushu and Hainan prefectures.
CONCLUSIONFrom 2001 to 2010, animal infected with plague was detected in successive years and human plague was very common in Qinghai. New infected animals and vector insects species and new epidemic areas were confirmed, hence the trend of plague prevalence for humans and animals is very active in Qinghai province.
Animals ; Antibodies, Bacterial ; blood ; China ; epidemiology ; Disease Vectors ; Humans ; Insect Vectors ; microbiology ; Plague ; epidemiology ; transmission ; Yersinia pestis ; classification ; isolation & purification
10.Catheter aspiration alone or combined with thrombolysis in the treatment of superior mesenteric artery embolism
Yuanquan HUANG ; Zhongzhi JIA ; Qi WANG ; Wenhua CHEN ; Zhongming HE ; Jun ZHANG ; Kai WANG ; Feng TIAN
Chinese Journal of Gastrointestinal Surgery 2014;(10):1018-1021
Objective To investigate the efficacy of catheter aspiration or combined with thrombolysis in the treatment of superior mesenteric artery embolism (SMAE). Methods Clinical and imaging data of 25 SMAE patients who underwent catheter aspiration or combined with urokinase thrombolysis in the First People′s Hospital and the Second People′s Hospital of Changzhou from January 2005 to July 2013 were retrospectively analyzed. Results Twenty patients were confirmed as SMA trunk embolism and 5 as SMA branch artery embolism. The embolic SMA trunks were completely recannulated by catheter aspiration in the above 20 cases, but small emboli embolized distal branch artery in 6 cases. These 6 patients plus above 5 patients with branch artery embolism received catheter aspiration combined with thrombolytic therapy. Among these 11 patients , complete open, partial open and non-open of branch arteries were found in 5, 3, 3 cases respectively, while collateral circulation increased significantly in non-open patients. During the follow-up period of (4.1±2.2) months, clinical symptom relief and digestive function recovery were observed in 24 cases. Only one case underwent bowel resection because of intestinal necrosis 24 hours after treatment and developed short bowel syndrome. Conclusion Catheter aspiration or combined with thrombolysis is a safe and effective method in treating SMAE.

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