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.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.
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.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.
6.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.
7.Electrocardiographically triggered CT angiography of the whole aorta and coronary arteries with high-pitch dual-source CT
Nan ZHANG ; Yu LI ; Yuan Lü ; Long ZHAO ; Wei GENG ; Zhanming FAN
Chinese Journal of Radiology 2013;(4):310-315
Objective To investigate the feasibility of comprehensive assessment of the whole aorta and coronary arteries (CA) simultaneously with high-pitch 128-slice dual-source CT ECG-gated FLASH protocol.Methods A total of 48 consecutive patients with suspected aortic diseases underwent CTA examination of the whole aorta and CA using a ECG-gated FLASH CT protocol (pitch =3.2) without heart rate (HR) control.Aorta,aortic valves and CA were shown with different post-processing modalities.The image quality of the aorta,aortic valves and CA was evaluated and compared according to HR (low HR group ≤ 65 bpm,high HR group > 65 bpm).The scan time,effective dose and contrast medium volume were recorded.Inter-observer differences were calculated by Kappa test.Differences between groups were analyzed by Mann-Whitney test with count data and variance test with measurement data.Results All examinations were completed successfully.The image quality was acceptable in the aorta,aortic valve (100%,48/48) and CA (94.0%,551/586).Percentage of diagnostic quality images was slightly higher in low HR group (94.7%,232/245,93.5%,319/341,Z =-2.504,P < 0.05).Inter-reader reproducibility was 95.8% (46/48) in the aorta and aortic valve,96.6% (566/586) in CA,yielded good agreement (Kappa =0.81 and 0.89).The mean attenuation of aorta and CA were higher than 300 HU,especially in low HR group (357.0-446.0) HU.Except proximal segment of right CA,the CNR of whole aorta and coronary arteries were significantly higher in the low HR group (24.5-29.0) than that in the high HR group (20.0-23.1,P <0.05).SNR of the whole aorta was higher in the low HR group (13.7-17.9) than the high HR group (11.5-13.9,P < 0.05).The mean scan time was (1.56 ± 0.08) s,effective dose was (4.12 ± 1.23) mSv (2.77-6.77 mSv),and contrast medium volume was (72.8 ± 2.1) ml.Conclusions CT angiography of whole aorta and coronary arteries could be performed simultaueously within 2 seconds with CT ECG-gated FLASH protocol.The images were of diagnostic quality for aortic and CA disease with low dose of radiation and contrast media.However,high HR could decrease the image quality of CA.
8.The expression of uPA and uPAR in patients with systemic inflammatory response syndrome
Li YU ; Ding LONG ; Xiaoling WU ; Junhui YANG ; Yuanchao ZHANG ; Feng GENG
Chinese Journal of Emergency Medicine 2011;20(10):1079-1083
Objective To test plasma levels of uPA,uPAR,D-dimer,IL-6 and TNF-α,and observe the relationships between uPA,uPAR and D-dimer,IL-6,TNF-αin patients with SIRS.Methods A prospective,clinical case-control study was adopted.Cases were collected from our hospital in January 2008 to January 2010,and all were > 55 years of age.Venous blood samples were collected via routine venipuncture.Eighty-five patients were divided into two groups according to diagnostic criteria of SIRS:SIRS group collected from Intensive Care Unit ( n =50) and non SIRS group collected from medical ward ( n =35).The control group comprised 30 unrelated healthy blood donors who visited the General Health Checkup Division at our hospital.Patients with (1) pregnant women; (2) cancer; (3) died after admitted into ICU in 7 days; (4) after cardiopulmonary resuscitation ; ( 5 ) with previous blood system diseases; (6) patients with SIRS before admitted into ICU were excluded from the study.uPA,uPAR,D-D,IL-6 and TNF-α in blood were detected by commercial enzyme-linked immunosorbent assay (ELISA) kits.The data was analyzed using SPSS version17.0.Data accorded with normal distribution of measurement was expressed as mean ± standard,and analyzed by independent-samples t test; non-normal distribution of measurement data,expressed by median,was analyzed with Mann-Whitney test.Relationships between plasma levels of uPA,uPAR and D-dimer,IL-6 TNF-α were analyzed using Spearman rank correlation test.To compare with blood level of uPA,uPAR,IL-6 and TNF-α in SIRS patients in the application of diagnostic value of MODS,we constructed the receiver operating characteristic curve ( ROC curve) for blood levels of uPA,uPAR,IL-6 and TNF-α in 24 h.Results The plasma levels of uPA,uPAR,D-dimer,IL-6 and TNF-αin patients of SIRS were obviously higher compared with non-SIRS and normal controls ( all P < 0.01 ).Correlation analysis showed that there was positive correlation between uPAR level and IL-6 level (r =0.395,P =0.004) ; there was positive correlation between uPAR level and TNF-αlevel ( r =0.606,P <0.01 ).There was no correlation between uPAR levd and D-dimer level ( r =- 0.069,P =0.632).There was no correlation between uPA level and D-dimer,IL-6 or TNF-α ( all p > 0.05).There ROC curve were made based on the abilities of uPAR,D-dimer,IL-6 and TNF-αlevels in 24 hours to diagnose MODS,and the ROC areas under the curves were 0.76,0.58,0.86,0.83 respectively.Conclusions These results demonstrate that uPA and uPAR play a major contributory role in patients with SIRS in the process of coagulation disorders,but the mechanism in SIRS is not the same.uPAR may play a central rolein the development of SIRS to MODS.
9.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 .
10.The investigation of glucocorticoid receptors in patients with polymyositis/dermatomyositis
Jiu-Hong LI ; Ning ZHAI ; Bo LI ; Gui-Jiao BI ; Long GENG ; Fang-Ji SONG ;
Chinese Journal of Rheumatology 2001;0(04):-
0.05).But GR number[sites/cell]and the expression of GR mRNA in PBMCs from PM/DM was significantly lower than those in healthy controls(P