2.Chemiluminescence in Whole blood and its Clinical application Ⅱ. Detection of phagocytic function of leucocytes from patients with various diseases
Chinese Journal of Immunology 1986;0(04):-
Measurement of whole blood CL was carried out to evaluate phagocytic of function of PMNL and opsonic activity of plasma from children (22 cases), adults (31), newborn infants (umbilical cord blood, 21), patients with bacterial infection (31), viral infections (21),and mild anemia (10). The following results were obtained: first, phagocytic function of PMNL and opsonic activity of plasma from normal children was compared to adults; second, phagocytic and/or metabolic activity of PMNL and opsonic ativity of plasma from newborn infants were lower then normal, third, PMNL of the patients with bacterial infections had been preactivated in vivo, their phagocytic function being enhanced, but, blood from the patients with viral infections showed almost no abnormalities; fourth, phagocytic activity of PMNL from the patients with mild anemia was similar to normal human. The significance of the results was also discused in the paper.
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.Postoperational Fungus Infection: An Analysis
Haiyan XU ; Haijun WANG ; Yingzhen LIU ; Yong GAO ; Qinglong XIAO
Chinese Journal of Nosocomiology 1994;0(04):-
OBJECTIVE To investigate the distribution of fungus infection and risk factor of postoperation patients with tumors. METHODS We analyzed 1 256 postoperation patients in our hospital ICU from Aug 2000 to Aug 2004,and found that there were 88 fungus infection patients(7%),the pathogens were tested and analyzed. RESULTS The fungus infection,which dominated in respiratory tract and digestive tact,had an increasing tendency,the most prevalent fungus of infection was Candida albicans. CONCLUSIONS The risk factors of fungus infection are mechanical ventilation,the useness of antibiotics,radiotherapychemotherapy,and invasive treatment,it is very important to diagnose early and treat in time.
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 Qinghuang Powder combined Chinese herbs for Pi-strengthening and Shen-reinforcing on HIF-1alpha in bone marrow mononuclear cells of myelodysplastic syndrome patients: an experimental research.
Fei GAO ; Yong-Gang XU ; Xiao-Hong YANG ; Rou MA
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(2):174-178
OBJECTIVETo study the effect of Qinghuang Powder (QHP) combined Chinese herbs for Pi-strengthening and Shen-reinforcing (CHPSSR) on hypoxia-inducible factor 1alpha (HIF-1alpha) in bone marrow mononuclear cells of myelodysplastic syndrome (MDS) patients.
METHODSChanges of HIF-1alpha in bone marrow mononuclear cells of MDS patients were detected in 25 MDS patients treated by QHP combined CHPSSR using flow cytometry. Meanwhile, 13 healthy subjects were recruited as the control group. Changes HIF-1alpha levels in various serial bone marrow mononuclear cells were detected.
RESULTS(1) Among the 25 patients in the treatment group, there were 19 patients effective and 6 patients ineffective, with the total effective rate being 76%. (2) Compared with before treatment, levels of peripheral blood WBC, Hb, PLT, and ANC significantly increased in the treatment group after treatment, showing statistical difference (P < 0.05, P < 0.01). (3) Compared with before treatment, the HIF-1alpha mean fluorescence intensity was enhanced in bone marrow lymphocytes, monocytes, granulocytes, and nucleated red blood cells of the treatment group after treatment (P < 0.05, P < 0.01). Compared with the control group, the HIF-1alpha mean fluorescence intensity was weakened in bone marrow lymphocytes, monocytes, and nucleated red blood cells of the treatment group before treatment; while it was obviously enhanced in granulocytes (P < 0.01). But after treatment the HIF-1alpha mean fluorescence intensity increased more in the granulocytes of the treatment group than in those of the control group (P < 0.01), but there was no statistical difference in bone marrow lymphocytes, monocytes, or nucleated red blood cells.
CONCLUSIONQHP combined CHPSSR could improve HIF-1alpha levels in bone marrow lymphocytes, monocytes, granulocytes, and nucleated red blood cells of MDS patients, thus improving Hb levels of MDS patients, and improving their anemia and correlated symptoms.
Adolescent ; Adult ; Aged ; Arsenicals ; therapeutic use ; Bone Marrow ; Bone Marrow Cells ; drug effects ; metabolism ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Middle Aged ; Monocytes ; drug effects ; metabolism ; Myelodysplastic Syndromes ; drug therapy ; metabolism ; Phytotherapy ; Young Adult
7.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.
8.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.
9.Effect of MOTOmed Intelligent Training System on Balance and Lower Limb Motor Function in Stroke Patients
Chunhua GAO ; Leyi XU ; Jie HUANG ; Feng XIAO
Chinese Journal of Rehabilitation Theory and Practice 2013;19(8):725-728
Objective To investigate the effect of MOTOmed intelligent training system training on balance and lower limb motor function in stroke patients. Methods 120 stroke patients were randomly divided into observation group (n=60) and control group (n=60) according to the random number table. Both groups were treated with routine rehabilitation training, the observation group received MOTOmed intelligent training system in addition. They were evaluated with Fugl-Meyer assessment (FMA), Functional Ambulation Category (FAC), Barthel index (BI), Motricity index (MI-L), modified Ashworth scale (MAS) and Berg balance scale (BBS) before and 4, 8, 12 weeks after treatment. Results There was no difference in the score of FMA, BI, MI-L, BBS, MAS, FAC and the maximum walking speed, stride length and stride frequency between 2 groups before treatment (P>0.05). The scores of FMA, BI, MI-L, BBS, FAC and the maximum walking speed stride length and stride frequency increased in the observation group and there was a uptrend 4 weeks, 8 weeks and 12 weeks after treatment (P<0.05). The score of MAS decreased in the observation group and there was a downtrend after treatment (P<0.05). All the indexes were better in the observation group than in the control group (P<0.05). Conclusion MOTOmed training system combined with routine rehabilitation training can improve the balance and lower limb motor function in stroke patients.
10.Construction of lentivirus vector containing human ?-catenin-EGFP and its expression in human hair follicle stem cells
peng-gao, YANG ; xiao-hui, HU ; feng-hou, GAO ; wei-rong, YU ; peng, XU ; yong, FANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(09):-
Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.