1.Variations of serum peptide spectrum in nephrotic syndrome patients
Weiguo SUI ; Yue ZHANG ; Jiejing CHEN ; Huaying LIU ; Guiying FEI ; Guimian ZOU ; Yong DAI
Journal of Chinese Physician 2011;13(6):760-764
Objective To compare the serum peptidome spectrum between nephrotic syndrome patients and normal controls, and to search for their variations. Methods The serum peptide profiling was determined by ClinProt magnetic bead enrichment and matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) in 17 mesangial proliferative glomerulonephritis (MsPGN) patients, 12 minimal change nephrotic syndrome (MCNS) patients, 10 membranous nephropathy (MN) patients, 10 focal segmental glomerulosclerosis (FSGS) patients, and 10 healthy volunteers. Results 5 differentially expressed polypeptides were screened out between MsPGN and normal controls (15.28±7.61, P<0.01). 7 differentially expressed polypeptides were screened out between MCNS and normal controls (2.16±1.59, P<0.01). 6 differential expressed polypeptides were screened out between MN and normal controls (35.48±13.71, P<0.01). 5 differential expressed polypeptides were screened out between FSGS and normal controls (18.06±8.07, P<0.05). The statistical significance was set at P<0.05. A Genetic Algorithm was used to set up the classification model between patients and normal controls. The model separated MsPGN, MCNS, MN and FSGS group from normal controls with a cross validation of 96.18%, 100%, 98.53% and 94.12%, respectively. The recognition capabilities were 100%. Conclusions The study established the serum peptidome spectrum for nephrotic syndrome by proteomic technology, and provided a new viewpoint to better understand the pathogenesis of nephrotic syndrome.
2.Intrinsic prokaryotic promoter activity of SUMO gene and its applications in the protein expression system of Escherichia coli.
Yanhong QI ; Zhurong ZOU ; Huaying ZOU ; Yunliu FAN ; Chunyi ZHANG
Chinese Journal of Biotechnology 2011;27(6):952-962
Nowadays, SUMO fusion system is important for recombinant protein production in Escherichia coli, yet a few aspects remain to be improved, including the efficacy for vector construction and protein solubility. In this study, we found the SUMO gene Smt3 (Sm) of Saccharomyces cerevisiae conferred an unexpected activity of constitutive prokaryotic promoter during its PCR cloning, and the gene coding regions of SUMOs in most species had a sigma70-dependent prokaryotic promoter embedded, through the prediction via the BPROM program developed by Softberry. By combining the characters of Sm promoter activity and the Stu I site (added at the 3'-terminal of Sm), and introducing a His-tag and a hyper-acidic solubility-enhancing tag, we further constructed a set of versatile vectors for gene cloning and expression on the basis of Sm'-LacZa fusion gene. Experimentally started from these vectors, several target genes were subcloned and expressed through blue-white screening and SDS-PAGE analysis. The results manifest a few of expectable advantages such as rapid vector construction, highly soluble protein expression and feasible co-expression of correlated proteins. Conclusively, our optimized SUMO fusion technology herein could confer a large potential in E. coli protein expression system, and the simultaneously established co-expression vector systems could also be very useful in studying the protein-protein interactions in vivo.
Amino Acid Sequence
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Base Sequence
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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genetics
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Lac Operon
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genetics
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Molecular Sequence Data
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Promoter Regions, Genetic
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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Saccharomyces cerevisiae Proteins
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biosynthesis
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genetics
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Small Ubiquitin-Related Modifier Proteins
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biosynthesis
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genetics
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Sumoylation
3.Investigation and analysis of fertility intention and psychological pressure of women of childbearing age
Chunyong ZHANG ; Huaying CHEN ; Yonghui ZHANG ; Chang′e ZOU
Journal of Chinese Physician 2022;24(11):1613-1615
Objective:To investigate the fertility intention and psychological pressure of women of childbearing age in Yanshan County, Jiangxi Province, and to provide data support for the targeted promotion of fertility.Methods:Using the method of random sampling, women of childbearing age in 10 communities in Yanshan County, Jiangxi Province were given questionnaires and psychological stress rating tables to understand the basic information, the status of marriage and childbearing, childbearing intentions, and the Chinese version of the Perceived Stress Scale (CPSS) score, and explore the related factors that may affect childbearing intentions and psychological stress of women of childbearing age.Results:In this survey, 1 500 questionnaires were distributed to women of childbearing age, 913 of which were returned, and 65.0%(593/913) of them were willing to have children. The CPSS score of women of childbearing age was 8-28(16.84±2.75). The CPSS scores of women of childbearing age in different occupations were statistically significant ( P<0.05). Public officials (including civil servants and employees of public institutions) had the least psychological pressure, while self-employed and other (unemployed) had the most psychological pressure. There was no significant difference in CPSS scores among women of childbearing age with different education levels, annual income levels, marital status and willingness to have children (all P>0.05). The correlation analysis showed that there was a statistical significance between the psychological pressure of women of childbearing age and their occupation ( r=0.230, P<0.05), and there was no significant correlation between other factors such as education level, annual income, fertility desire, fertility status, etc. and psychological pressure (all P>0.05). Conclusions:The fertility intention desire of women of childbearing age in Yanshan County, Jiangxi Province is basically normal, and the desire to have more children is not strong. There is no significant correlation between fertility intention and psychological pressure. The psychological pressure of women of childbearing age is significantly related to their occupation.
4.Applied anatomical study and clinical application of the caudate lobe boundary and ductal system of the liver
Yihua HUANG ; Xinhua JIANG ; Hang YUAN ; Huaying ZOU ; Wei MAO
Chinese Journal of Hepatology 2022;30(10):1100-1106
Objective:To explore the relationship between the hepatic caudate lobe boundary and the ductal system so as to guide the identification of the anatomical relationship during liver surgery.Methods:The specific parts were observed and the liver parenchyma was removed according to 41 cadaveric liver autopsy specimens. The critical relationship between the hepatic caudate lobe and other ducts was observed to explore the reticular duct structure.Results:The plane formed by the hepatic hilar plate and Arantius ligament served as the boundary between the caudate lobe and other hepatic lobes. The caudate lobe hepatic portal vein was composed of numerous small branches from its left and right branches. The portal vein adjacent to the vena cava was mainly derived from the left branch, and to a lesser extent from the right branch. Blood was drained straight from the caudate lobe vein into the inferior vena cava via the short hepatic vein. There were three or four bile duct branches in the caudate lobe. The main source of arterial blood flow were the left and right branches of the hepatic artery. An avascular zone of loose connective tissue was found between the caudate lobe and the retrohepatic inferior vena cava.Conclusion:The hepatic caudate lobe is an independent lobe. During hepatic caudate lobe surgery, the plane formed by the hepatic hilar plate and Arantius ligament can serve as the boundary between the caudate lobe and other hepatic lobes and be used for anatomical site identification. The duct system of the caudate lobe's is complicated, but it also has its own distinct regularity.
5.The water-soluble TF3 component from Eupolyphaga sinensis Walker promotes tibial fracture healing in rats by promoting osteoblast proliferation and angiogenesis
Binghao Shao ; Xing Chen ; Jin' ; ge Du ; Shuang Zou ; Zhaolong Chen ; Jing Wang ; Huaying Jiang ; Ruifang Lu ; Wenlan Wang ; Chunmei Wang
Journal of Traditional Chinese Medical Sciences 2024;11(2):245-254
Objective:
To determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability.
Methods:
A modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels.
Results:
Tu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment.
Conclusion
TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.