1.Screening of binding proteins of HMGB1 promoter by phage display technique
Ning DING ; Hui XIAO ; Ju GAO ; Lixin XU ; Shouzhang SHE
Chinese Journal of Pathophysiology 2010;26(1):28-31
AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.
2.ERK signal is pathway involved in mechanical stretch induced HMGB1 expression in alveolar epithelial cells
Ning DING ; Hui XIAO ; Ju GAO ; Lixin XU ; Shouzhang SHE
Acta Anatomica Sinica 2009;40(6):919-922
Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.
3.p38 signaling pathway involved in the expression of HMGB1 in a rat model of ventilator-indnced lung injury
Ning DING ; Hui XIAO ; Ju GAO ; Lixin XU ; Shouzhang SHE
Chinese Journal of Emergency Medicine 2009;18(11):1169-1172
Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.
4.Effect of Peitu Shengjin Recipe on Nutritional States and Immune Functions of Stable Phase COPD Patients.
Jing GONG ; Ning CHEN ; Xiao-mei HAO ; Hui LI
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(5):534-536
OBJECTIVETo explore the effect of Peitu Shengjin Recipe (PSR) on nutritional states and immune functions of stable phase chronic obstructive pulmonary disease (COPD) patients.
METHODSTotally 62 stable phase COPD patients were randomly assigned to the treatment group (30 cases) and the control group (32 cases). All patients inhaled Seretide (50/500 µg), twice per day. Besides, patients in the treatment group additionally received PSR, one dose per day. After three months of treatment, the COPD assessment test (CAT) score, the index of nutritional states [including body mass index (BMI) , thickness of skin fold (TSF), mid-arm muscle circumference (MAMC), serum albumin, serum prealbumin], and immune functions (including IgA, IgM, and IgG) were compared between the two groups.
RESULTSBy the end of the treatment, the CAT score decreased more obviously in the treatment group than in the control group (P < 0.05). The improvement of BMI, TSF, MAMC, serum albumin, and serum prealbumin was better in the treatment group than in the control group (P < 0.05). IgM and IgG also increased more in the treatment group (P < 0.05). There was no statistical difference in IgA between the two groups (P > 0.05).
CONCLUSIONSAdditionally use of PSR could improve nutritional states and immune functions of stable phase COPD patients to some extent. Meanwhile, it also could improve their health related quality of life.
Body Mass Index ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Nutritional Status ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; immunology ; Quality of Life ; Serum Albumin
5.Construction of red fluorescent protein reporter gene vector containing HMGB1 promotor and its activity under mechanical stretch
Ning DING ; Hui XIAO ; Ju GAO ; Lixin XU ; Shouzhang SHE
Journal of Third Military Medical University 2003;0(22):-
Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.
6.Effect of hydroxyethyl starch 130/0.4 on ICAM-1 expression in endotoxin-induced acute lung injury in rats and role of MAPK signal pathway in its expression
Ning DING ; Hui XIAO ; Lixin XU ; Shouzhang SHE
Chinese Pharmacological Bulletin 1987;0(03):-
Aim To examine the effect of different dosages hydroxyethyl starch(HES)130/0.4 on intercellular adhesion molecule 1(ICAM-1)expression in lung tissue of acute lung injury in endotoxemic rats and explore the role of MAPKs pathway in its expression.Methods Thirty six healthy Sprague Dawley(SD)rats weighing 270~320 g were randomly divided into 6 groups with 6 animals in each group.In group H1-H4,1 min after lipopolysaccharide(LPS)5 mg?kg-1 intravenously administration,HES 130/0.4 with 3.75,7.5,15,30 ml?kg-1 were infused intravenously respectively at a rate of 0.2 ml?min-1.In group L,saline instead of HES 130/0.4 was administered.Group N served as control by giving the same volume of saline.The animals were anesthetized with pentobarbital.Right external jugular vein was injected with LPS.The animals were killed 4 hours after LPS injection for determination of total protein,WBC,MPO,W/D,ICAM-1 protein and mRNA and MAPKs activity.Results Compared with control group,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were increased significantly in group L.Compared with group L,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were decreased significantly in group H1 and H2,especially at the dosage of 7.5 ml?kg-1.Conclusion HES 130/0.4(7.5 ml?kg-1)can attenuate inflammatory response of acute lung injury induced by LPS,which may be related with inhibiting the expression of ICAM-1 protein and mRNA through MAPKs signal pathway.
7.Epidemiological characteristics of canine Echinococcus infection in Qinghai⁃ Tibet Plateau of China
Hui LIU ; Ning XIAO ; Shijie YANG ; Dong WANG ; Jia PENG
Chinese Journal of Schistosomiasis Control 2017;29(2):129-138
The Qinghai⁃Tibet Plateau is known as one of the highest endemic areas of echinococcosis However,the dog infection rates of Echinococcus granulosus in the plateau regions were similar to other non⁃Tibetan areas with the high endemic,and most of the rates were below 40%. The infected dogs with E. multilocularis were ubiquitous in Ganzi Prefecture of Sichuan Province and Qinghai Province where many survey data were done and available,which was much different from those in non⁃Ti⁃betan areas where the geographical distribution of dogs infected with E. multilocularis was sporadic. The total infection rates of Echinococcus in dogs kept stable from 1983 to 2009 in Ganzi Prefecture of Sichuan Province and did not show much variation from 2000 to 2014 in Qinghai Province as well. Since 2006,the national comprehensive prevention and control strategy and mea⁃
sures against echinococcosis have been launched in China,and significant progress has been made. In the endemic Tibetan area of Sichuan,the general Echinococcus infection rates in dogs were 28.10%,15.87%,19.22%,3.28% and 1.11% from 2009 to 2013,respectively,and the Echinococcus coproantigen⁃positive rate in Gannan Prefecture of Gansu Province and parts of Qing⁃
hai Province also decreased. This paper reviews the literature on the characteristics of dog infections in the Qinghai⁃Tibetan Plateau,so as to provide useful information to support echinococcosis control and prevention there.
8.Clinical Analysis of Cyanotic Congenital Heart Diseases in 102 Newborns
ning, ZHAO ; wei, ZHOU ; hui, LV ; xiao, RONG
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To analyze the distribution and clinical characters of cyanotic congenital heart diseases in newborns.Method We examined the suspicious cases with color doppler ultrasonic cardiogram(CFM),and compared with the results of autopsies.Results The most common congenital diseases in the group were complete transposition of the great arteries,with the proportion of 34.4%, and most of them were compounded cardiovascular disorders. The most common symptom was cyanosis, took the proportion of 84.3%; some patients had no cyanosis, tachypnea and murmurs.The cases with cardiovascular disorders most were combined with type Ⅱrespiratory failure and metabolic acidosis.Conclusions There are many types of congenital heart diseases, the symptoms are untypical,so the suspicious cases shall examine with CFM as early as possible.
9.Expression of Thyroid Hormone Receptor Interacting Protein 3 in Children with Acute Lymphocytic Leukemia and Its Significance
xiao-hui, LI ; jian-bai, ZHANG ; fang, NING ; jing, LI
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To study the molecular mechanism of the remission in children with acute lymphocyte leukemia(ALL),as well as the expression of thyroid hormone receptor interacting protein 3(TRIP3) gene in children with ALL and explore the relationship between TRIP3 and ALL.Methods Fasting venous blood 2-4 mL was collected,anticoagulanted with ethylenediamine tetraacetic acid(EDTA),then perpheral blood mononuclear cells was collected by Ficoll density gradient centrifugation,total RNA was extracted by Trizol one step method.Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect TRIP3 expression in peripheral blood lymphocytes in 73 ALL children of different stages and 20 normal children.The relationship between TRIR3 expression in children and ALL release was analyzed.Results 1.Expression of TRIP3 was significantly lower both after initial treatment and during recrudescence than that in normal children(Pa0.05).3.In children with ALL after initial treatment,the remission rate was significantly higher in TRIP3 positive patients than in TRIP3 negative ones(remission rate discern 25.0% vs 84.2%,P
10.Application and expectation of robot-assisted operation system in gastrointestinal surgery.
Chinese Journal of Gastrointestinal Surgery 2012;15(8):778-780
With the development of minimally invasive surgery in China, the da Vinci surgical system was used more and more widely in gastrointestinal surgery. This paper summarizes the development history of the artificial intelligence-assisted surgery system, the application and in the promotion of gastrointestinal surgery in China. Application of this new technology promotes further the innovation and development of minimally invasive surgery, and will drive leap in the field of gastrointestinal minimally invasive surgery.
Digestive System Surgical Procedures
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methods
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Humans
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Minimally Invasive Surgical Procedures
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Robotics
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Surgery, Computer-Assisted