2.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.
3.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.
4.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.
5.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.
6.A comparative study of HPV infection in condyloma acuminate tissues of vulva,vagina and cervix
Huimin SHAN ; Jianxiang GENG ; Jing MEI ; Xiurong LONG ; Lin XIA ; Hongjing WANG ; Xue ZHAO
International Journal of Laboratory Medicine 2016;37(16):2239-2241
Objective To study the distribution status and clinical significance of human papillomavirus(HPV) infection geno‐types in condyloma acuminate(CA) tissues of vulva ,vagina and cervix .Methods The gene‐chips combined with polymerase chain reaction (PCR) technology were utilized for detecting 23 kinds of HPV genotypes in tissue specimens from 63 cases of vulval CA , 61 cases of vaginal CA and 65 cases of cervical CA .Their clinical pathological data were analyzed .Results In 63 cases of vulval CA ,56 cases were HPV positive with the HPV infection rate of 88 .89% (56/63) ,in 61 cases of vaginal CA ,55 cases were HPV positive with the HPV infection rate of 90 .16% (55/61) ,and in 65 cases of cervical CA ,62 cases were HPV positive with the HPV infection rate of 95 .39% (62/65) .Conclusion HPV infection is closely related to the CA pathgenesis in vulva ,vagina and cervix . HPV6 and HPV 11 are main stream genotypes ,in which vulval CA is most common .The gene‐chips combined with PCR technology is a method suitable for HPV typing diagnosis ,and has the characteristics of good sensitivity and high specificity ,which has an im‐portant significance for clinical diagnosis ,treatment and vaccine study of CA in femal vulva ,vagina and cervix .
7.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.
8.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.
9.The research of that Shikonin effects on VEGF production in IL-17-stimulated HaCaT cells
Min HANG ; Long GENG ; Hongwei REN ; Huiming QU ; Xue WANG ; Yongzhi JI ; Zhongxiang WEI ; Hongbo ZHOU
Chinese Journal of Microbiology and Immunology 2011;31(8):685-688
Objective To investigate whether IL-17 could stimulate the vascular endothelial growth factor (VEGF) production on HaCaT cells alone. We also investigated whether shikonin could inhibited the proinflamation effects of interleukin-17(IL-17) acting on HaCaT cells. MethodsWe examined the expression of VEGF by double antibody sandwich enzyme-linked immunosorbent assay ( ELISA ) and realtime polymerase chain reaction(RT-PCR) in HaCaT cells and the cell supernatant. The viability of HaCaT cells in the drug group was detected by the Cell Counting Kit-8 (CCK-8). ResultsThe expression of VEGF in different time IL-17-stimulated groups on HaCaT cells and the cell supernatant were higher than the control group( P<0.001 ). The expression of VEGF in different drug treatment groups on HaCaT cells and the cell supematant were lower than the stimulated group by IL-17 ( P<0. 001 ). The cell viability of different drug treatment groups have no significant difference( P>0.05 ). ConclusionWe show that IL-17 specifically and time-dependently augmented and induced VEGF expression on HaCaT cells and the cell supernatantThen shikonin markedly inhibited the increase tengency of IL-17 effection on HaCaT cells and the cell supematant level.
10.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.