2.Proliferation of desmoglein 3-stimulated T lymphocytes from patients with pemphigus vulgaris
Jie HU ; Zhongxiang WEI ; Yi JIANG ; Zaifu LIANG ; Long GENG
Chinese Journal of Dermatology 2010;43(7):474-477
Objective To investigate the impact of desmoglein 3 (Dsg3) on the proliferation of peripheral T lymphocytes from patients with pemphigus vulgaris (PV).Methods Peripheral blood mononuclear cells (PBMCs) were obtained from 12 patients with PV and 22 normal human controls,cultured with or without the presence of Dsg3 or phytohemagglutinin for 3 days.Flow cytometry was performed to detect the changes of T-lymphocyte subsets in PBMCs stimulated with Dsg3 and proliferation of T-lymphocytes.Results In patients with PV,the percentage of Th2 and Th1 cells was 12.17%±5.32% and 4.08%±1.50%,respectively in Dsg3stimulated PBMCs,9.84%±5.41% and 3.91%±1.38%,respectively in non-stimulated PBMCs.Increased percentage of Th2 cells was observed in Dsg3-stimulated and non-stimulated PBMCs from patients with PV compared with those from normal human controls (both P<0.05).After stimulation with Dsg3,there was a significant proliferation of T cells from patients,and the proliferation rate of CD4+T cells was 4.65%±3.28%,which was significantly higher than that from normal controls(P<0.05).Conclusion Dsg3 can induce the specific proliferation of CD4+T cells,especially Th2 type CD+4 T cells,from patients with PV.
3.Study of association between HLA class Ⅱ haplotypes and pemphigus vulgaris in Han nation of northeast China
Long GENG ; Ning ZHAI ; Xiuping HAN ; Hongxi NIU ; Fangji SONG
Chinese Journal of Immunology 1985;0(05):-
Objective:To investigate the relationship between pemphigus vulgaris(PV) and HLA-DR,DQ haplotypes in Han nations of northeast China.Methods:Polymerase chain reaction-sequence specific primers(PCR-SSP) method was used to detect the HLA-DRB1 and DQB1 alleles of 27 PV patients of Han nation of northeast China, analysed haplotyes and compared with 99 healthy controls.Results:Compared with control group, the frequencies of the haplotypes of HLA-DRB1*140x-DQB1*0503,DRB1*140x-DQB1*0201,DRB1*120x-DQB1*0503 and DRB1*140x-DQB1*0302 increased significantly in PV group. After statistical test, the difference between the two groups was significant.Conclusion:The special haplotypes may contribute to genetic susceptibility to PV in northeast Chinese.
4.Effect of hydrogen on postoperative cognitive function in aged mice
Hongfang GENG ; Long HE ; Jiaqiang ZHANG ; Chenxi LI ; Fanmin MENG
Chinese Journal of Anesthesiology 2017;37(5):581-584
Objective To evaluate the effect of hydrogen on the postoperative cognitive function in aged mice.Methods Sixty aged male Kunming mice,aged 12-14 months,weighing 35-45 g,were assigned into 3 groups (n=20 each) using a random number table:sham operation group (group Sham),partial hepatectomy group (group PH) and partial hepatectomy plus hydrogen-enriched saline group (group PH-H).On preoperative day 3,during operation and on postoperative day 3,hydrogen-enriched saline 10 ml/kg was intraperitoneally injected once a day in group PH-H,and the equal volume of normal saline was intraperitoneally injected once a day in S and PH groups.The spatial probe test was performed on postoperative day 7 to evaluate the cognitive function.Ten mice were selected on postoperative days 3 and 7 (after the end of the spatial probe test),and blood samples were collected for determination of serum tumor necrosis factor-alpha (TNF-α),interleukin-lbeta (IL-1β) and high-mobility group box 1 protein (HMGB1) concentrations by enzyme-linked immunosorbent assay.The animals were sacrificed after blood sampling,brains were removed and hippocampi were isolated for detection of the expression of TNF-α,IL-1β and HMGB1 by Western blot.Results Compared with group S,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly decreased,the serum concentrations of TNF-α,IL-1β and HMGB1 were increased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was up-regulated in group PH (P<0.05),and no significant change was found in the variables mentioned above in group PH-H (P>0.05).Compared with group PH,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly increased,the serum concentrations of TNF-α,IL-1β and HMGB1 were decreased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was down-regulated in group PH-H (P<0.05).Conclusion Hydrogen can improve the postoperative cognitive function in aged inice.
5.The Selection of Thrombin-specific scAbs by Alternative Panning Strategy
Long HONG ; Bai-Cheng YANG ; Sheng-Geng ZHU ;
Microbiology 1992;0(03):-
Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.
6.Study on Dsg3 induced activation of T cells in first degree relatives of patients with pemphigus vulparis
Zhongxiang WEI ; Long GENG ; Jie HU ; Zhengrong SUN ; Shuyan DU ; Yi JIANG
Chinese Journal of Microbiology and Immunology 2009;29(12):1090-1093
Objective To investigate The Th1/Th2 and Tc1/Tc2 polarization in the peripheral blood of first degree relatives of pemphigus vulgaris(PV) and healthy control individuals, and to approach the mechanism of the Dsg3-specific autoimmunity in PV. Methods The peripheral blood mononuclear cells (PBMC) from first degree relatives and healthy control was stimulated for72 h with Dsg3 and without Dsg3. Th1/Th2, Tc1/Tc2 was assessed by four-color flow cytometry. Results The mean frequency of Dsg3-spe-cific Th2 cells for PV antibody positive first degree relatives was 10.13%±3.72%, compared with stimula-tion without antigen 7.28%±3.58%, the difference was significant (P<0.05). The percentage Dsg3-spe-cific Th2 was markedly higher in the PV antibody positive first degree relatives group than that in the control group(10.13%±3.72% vs 6.10%±2.82%, P<0.05) , Tc2 was markedly higher also (20.01%± 10.43% v514.91%±8.06%, 20.01%±10.43% vs 9.58%±5.49%, P<0.05). Conclusion When Dsg3 stimulated PBMC were used to stimulate autologous T cells an increased amount of Th2 and Tc2 was observed, it is implied that the imbalance of Th1/Th2, Tc1/Tc2 might play an important role in the initia-tion of PV.
7.Preparation of peri-epiphyseal fracture models in young rabbits:cross-epiphyseal plate internal fixation affects epiphyseal growth?
Qingda CUI ; Wei LIU ; Xin WANG ; Long MA ; Shiqian ZHANG ; Qi PAN ; Zhenggang BI ; Shuo GENG
Chinese Journal of Tissue Engineering Research 2015;(44):7148-7155
BACKGROUND:There are various therapies for children limb fractures involving the epiphysis or the metaphysis. According to the different methods, studies on the growth of the epiphyseal plate are a lot, most of which focus on the effects of Kirschner wires with different diameters or holow screw internal fixation on the development of epiphyseal plate. However, there are rare studies on the influence of cross-epiphyseal plate internal fixation on the growth of epiphyseal plate as wel as the influence level. OBJECTIVE:To prepare a peri-epiphyseal fracture model in young rabbits and to observe the effects of cross-epiphyseal plate implantation and removal on the growth of epiphyseal plate. METHODS: Traverse fracture models were made 5 mm above the right femoral distal epiphyseal plate of 60 young rabbits, and then fixed with suitable “L” steel plate and four screws across the epiphyseal plate and peri-epiphyseal fracture line. The left side served as control. Eight rabbits were kiled and observed at 2, 4, 8, 12 weeks after modeling, respectively, to take out the femoral specimens for measurement of femoral length, thickness of the epiphyseal plate, and number of mastocytes per unit column. Histopathology observation was done and changes in mastocytes and thickness of the epiphyseal plate were detected. Another seven rabbits were selected to remove the metal plate, continued to feed for 2 weeks and finaly executed to observe the above-mentioned indexes. RESULTS AND CONCLUSION: (1) There were significant differences in the above indexes between the plate and control groups at 4, 8, 12 weeks after modeling (P < 0.05 orP< 0.001) but not at 2 weeks after modeling (P> 0.05). These findings indicate that within 2 weeks after cross-epiphyseal plate internal fixation, proper pressue has no remarkable influence on the growth of epiphyseal plate; but after persistent internal fixation (> 4 weeks), the growth of epiphyseal plate can be partialy or completed retarded. (2) At 2 and 4 weeks after modeling, the plate was removed, and 2 weeks later, the femoral length, thickness of the epiphyseal plate and mastocyte counting per unit column were improved to different extents, and there were no differences between the plate and control group (P > 0.05). At 8 and 12 weeks after modeling, the plate was removed, and 2 weeks later, the femoral length and thickness of the epiphyseal plate were shortened, and the number of mastocytes per unit column was decreased obviously, which significantly differed from the control group (P < 0.001). These findings indicate that the chondrocytes in the proliferative and hypertrophy layers lose the differentiation and proliferation abilities, and the femoral length and epiphyseal plate thickness are difficult to recover.
8.Mechanical and biological properties of porous calcium phosphate cement/fibrin glue for bone regeneration in vitro
Jingjing DONG ; Long BI ; Na LI ; Yu GUO ; Qiang XU ; Wei LEI ; Geng CUI
Chinese Journal of Tissue Engineering Research 2014;(30):4831-4837
BACKGROUND:Fibrin glue introduced into calcium phosphate cement has not been confirmed whether this way could overcome the compressive limits and the low degradation of calcium phosphate cement and to modify the biological properties of calcium phosphate cement. OBJECTIVE:To explore the mechanical and biological properties of calcium phosphate cement/fibrin glue at different powder/liquid ratio for bone regeneration in vitro. METHODS:Calcium phosphate cement and fibrin glue were mixed at ratios of 1:1, 3:1, 5:1 (mL/g), and the pure calcium phosphate cement served as controls. Setting time, scanning electron microscope and the biomechanical test were used to analyze the composite scaffold structure, physical performance and the mechanical properties. Passage 3 osteoblasts were respectively inoculated on the material surface of the four groups, and pure cells served as blank controls. celladhesion, proliferation and alkaline phosphatase activity were observed. RESULTS AND CONCLUSION:The initial and final setting time of calcium phosphate cement/fibrin glue at 1:1 and 3:1 (mL/g) was higher than that in the control group (P<0.05), while the initial and final setting time of calcium phosphate cement/fibrin glue at 5:1 (mL/g) was lower than that of the control group (P<0.05). Scanning electron microscope showed smoother and denser surface of composite scaffolds compared with the pure calcium phosphate cement. The aperture of the composite scaffolds was decreased with the increasing concentration of fibrin glue. The compressive strength of composite scaffolds at 3:1 and 5:1 was higher than that of the control group (P<0.05), while the modulus of the composite scaffolds at 1:1, 3:1, 5:1 was higher than that of the control group (P<0.05). celladhesion, proliferation and alkaline phosphatase activity showed no difference among the three composite scaffold and control groups, but al higher than the blank control group (P<0.05). These findings indicate that fibrin glue introduced into calcium phosphate cement can overcome the low-strength limits of calcium phosphate cement, and maintain the good biological properties of calcium phosphate cement for bone regeneration.
9.Comparative study on fluorescent quantitation polymerase chain reaction and gene-chips typing method in genotyping HPV
Xiurong LONG ; Jianyun LAN ; Jianxiang GENG ; Xuemei FAN ; Lin XIA ; Hongjing WANG ; Jing MEI ; Xue ZHAO
International Journal of Laboratory Medicine 2015;(23):3385-3387
Objective To compare the sensitivity of fluorescent quantitation polymerase chain reaction (fluorescent quantitation method) and gene‐chips typing method(gene‐chips method) in the detection of human papillomavirus(HPV) ,and to analyse differ‐ences and clinical significance .Methods A total of 246 women were selected as subjects ,among them ,111 cases of cervical exfolia‐ted cells and 135 cases of cervical tissues were collected and detected .15 kinds of high‐risk HPV genetypes were detected in all sub‐jects by using fluorescent quantitation method and gene‐chips method respectively ,and the detection results were compared . Results The sensitivity of the fluorescent quantitation method in detecting HPV was 55 .28% and that of the gene‐chips method was 55 .69% ,there was no statistically significant difference in sensitivity between the two methods (P>0 .05) .The two methods had relative high conformance(κ=0 .745) .The positive rate of HPV infection was increased with the progression of cervical dis‐ease .Conclusion The fluorescent quantitation method and the gene‐chips method have a relative high conformance ,and both with high sensitivity in detecting HPV .The severity degree of cervical cytological and histological changes may be positively correlated with HPV infection .
10.Clinical evaluation of linezolid in 15 cases of severe lung infections caused by methicillin-resistant Staphylococcus aureus
Tao XU ; Li YU ; Junhui YANG ; Ding LONG ; Yuanchao ZHANG ; Feng GENG
Chinese Journal of Infection and Chemotherapy 2009;09(4):264-266
Objective To evaluate the efficacy and safety of linezolid in the treatment of severe lung infections caused by methicillin-resistant Staphylococcus aureus (MRSA).Methods Fifteen patients admitted to ICU due to severe lung infection caused by MRSA received linezolid treatment. WBC, lactic acid (LAC), IL-1β, IL-6, and TNF-α levels were measured before and after treatment. Results Clinical efficacy rate was 73.3%. The level of WBC, LAC and inflammatory cytokines decreased significantly after linezolid treatment (P<0.01).Conclusions Linezolid shows good efficacy in the treatment of severe lung infections caused by MRSA.