Objective To investigate the synergistic effect of calcium channel blocker on cyclosporine-induced gingival overgrowth (GO). Methods 130 renal transplant patients treated with cyclosporine were divided into group A (with calcium channel blocker) and group B (without calcium channel blocker). Demographic, pharmacologic and periodontal data were recorded. The prevalence and severity of GO were compared between the two groups. Three calcium channel blockers, including nifedipine, amlodipine and felodipine, were administered in the patients of group A. The relationships between these three calcium channel blockers and the prevalence of GO were analyzed. Results The patients receiving calcium channel blocker showed significantly higher prevalence of GO (44/73,60 % ) than those without calcium channel blocker (22/57, 39 %) (P＜0. 05). A higher proportion of mild GO (37 %) in group A was also observed than in group B (19 %, P＜0. 05). There were no significant differences in the proportions of moderate and/or severe GO between the two groups (P＞0. 05). Periodontal variables, including plaque index and papilla bleeding index, were significantly higher in GO patients than in those without GO in both two groups (P＜0. 05). In addition, the prevalence of GO in patients receiving nifedipine (77 %) was higher than in those receiving amlodipine (57 %) or felodipine (50 %). Conclusion The combination with calcium channel blocker is a risk factor of cyclosporine-induced GO and the use of nifedipine should be avoided for these at-risk patients.

Objective To establish a novel method to detect autoantibodies against platelatespecific receptors by flow cytometric bead assay and study its clinical application. Methods The beads were coated with monoclonal antibodies SZ2, SZ22, SZ21 and 7E3 against platelet GP Ⅰ b, GP Ⅱ b, GP Ⅲa and GP Ⅱ b/Ⅲ a, respectively. Captured platelet glycoprotein and beads complex was detected by FITC labeled polyclonal goat antihuman immunoglobulin using flow cytometer. The platelet samples that reacted with antibodies (SZ2, SZ22, SZ21 and 7E3) negatively and positively were tested, respectively. Each sample was repeated 20 times to generate intra-day CV for the MFI and once a day for 8 days to generate inter-day CV values. The 85 ITP patients, 17 NITP patients and 50 controls from the First Affiliated Hospital of Soochow University during March 2006 to December 2008 were included in the studies. The sensitivity and specificity of these four platelet antibodies to diagnose ITP were analyzed using ROC curve. The results were compared with MAIPA. Results The CV of the intra-day-assay for samples negative to antibody SZ2, SZ22,SZ21 and 7E3 were 3.26%, 2. 86%, 1.65% and 4. 94%, respectively; While the CV of the intra-day-assay for samples positive to antibody SZ2, SZ22, SZ21 and 7E3 were 6. 16%, 4. 88%, 5.20% and 5. 85%,respectively. The CV of the inter-day-assay for samples negative to antibody SZ2, SZ22, SZ21 and 7E3 were 5. 86%, 4. 74%, 5.69% and 7.56%, respectively; While the CV of the inter-day-assay for samples positive to antibody SZ2, SZ22, SZ21 and 7E3 were 7.53%, 5.49%, 7.11% and 6.25%,respectively. The MFI for SZ2 in ITP group, NITP group and healthy control group were 1.49(0. 88-16. 24),1.12(1.00-1.33), 1.01 (0. 83-1.37), respectively, which showed significant differences (H = 36.89,P＜0.01). The MFI for SZ22 in the three groups were 1.55 (0.84-11.30), 1.13(1.03-1.29), 0.98(0. 85-1.24), respectively (H=28.41, P ＜0.01). The MFI of SZ21 were 1.50 (0.87-11.04), 1.13(0.97-1.32), 1.05 (0.85-1.48), respectively (H=54.42, P＜0. 01). The MFI for7E3 were 1.51(0. 84-9.81), 1.05(0.86-1.13), 1.03 (0.74-1.28), respectively (H =31.97, P ＜0.01). Based on ROC analysis, with cut-off values of 1.37, 1. 24, 1.48 and 1.28 for SZ2, SZ22, SZ21 and 7E3,respectively, the AUC were 0. 86, 0.90, 0. 87 and 0. 84, respectively. The sensitivities of the assays were 58. 82% (50/85), 52. 94% (45/85), 52.94% (45/85) and 51.76% (44/85), respectively. When all four antibodies were used, the sensitivity was increased to 74. 12% (63/85), which was higher than that of MAIPA [ 50. 59% (43/85) ,χ2 = 6. 78, P ＜ 0. 05) ]. Conclusion Flow cytometric bead assay can be used to detect four platelet-specific autoantibodies simultaneously, and may be a useful method to aid in the diagnosis of ITP.

AIM: To investigate the clinical significance in determination of the P- selectin levels in subjects with prethrombotic state or thrombosis by flow cytometry (FCM). METHODS: The P- selectin expression on platelet membrane in 42 patients with diabetes mellitus, 33 with hyperlipidemia, 23 with cerebral infarction and 20 healthy individuals, were analyzed using fluorescently - labeled SZ - 51 by direct FCM comparing with indirect FCM and enzyme- linked immunosorbent assay (ELISA). RESULTS: The level of P- selectin on platelet membrane is higher in DM (23.92 % + 15.83 % ), in hyperlipidemia ( 18.34 % + 9.46 % ) and in cerebral infarction ( 19.32 % + 10.38 % ) than normal subjects (3.38 % + 1.11% ) ( P < 0.01 ). In addition, similar results on P - selectin were obtained by indirect FCM and ELISA in patients with DM and cerebral infarction. CONCLUSION: FITC - labeled SZ - 51 - IgG can be used in FCM, and it would be a new and sensitive method in detecting platelet activation.

AIM: To investigate the clinical significance in determination of the P-selectin levels in subjects with prethrombotic state or thrombosis by flow cytometry (FCM). METHODS: The P-selectin expression on platelet membrane in 42 patients with diabetes mellitus, 33 with hyperlipidemia, 23 with cerebral infarction and 20 healthy individuals, were analyzed using fluorescently-labeled SZ-51 by direct FCM comparing with indirect FCM and enzyme-linked immunosorbent assay (ELISA). RESULTS: The level of P-selectin on platelet membrane is higher in DM (23.92%?15.83%), in hyperlipidemia (18.34%?9.46%) and in cerebral infarction (19.32%?10.38%) than normal subjects (3.38%?1.11%) (P

AIM: To study the clinical significance of determining serum anti-endothelial cell antibodies (AECA) and thrombopoietin (TPO) in the differential diagnosis of idiopathic thrombocytopenic purpura (ITP) and systemic lupus erythematosus (SLE). METHODS: Serum AECA and TPO in 76 patients with ITP, 41 patients with SLE and 50 normal individuals were detected by ELISA method. RESULTS: Serum AECA level in SLE and ITP was much higher than that in normal group (P0.05), but serum TPO level in SLE was much higher than that in ITP and normal groups (P

Objective:To study the amino acid and codon usage profile of HIV-1 Env gene in donors whose serum exhibit highly broad cross-neutralizing activity. Methods:The samples were divided into highly broad cross-neutralizing activity group (hBCN + group) and non-highly broad cross-neutralizing activity group (hBCN - group) based on whether the neutralization breadth was higher than 90% or not. Full-length Env genes were amplified by single genome amplification (SGA) method from patients′ plasma samples, and the characteristics of Env sequences in hBCN + group were compared with hBCN - group. The correspondence analysis (COA) on relative amino acid usage (RAAU), adaptability to host based on similarity index D( A, B) and relative synonymous codon usage (RSCU) values of Env genes (hBCN + and hBCN -) with respect to human host RSCU were analyzed. Results:Correspondence analysis showed that the RAAU data of hBCN + group and hBCN - group were distributed along the two main axes to form two relatively separated clusters, indicating that the Env genes of the two groups had relatively unique amino acid usage patterns; the similarity index calculation results showed that hBCN + group (0.097) was lower than the hBCN - group (0.102), in addition, the Env gene of the hBCN + group had less frequency of similarly selected codons with human host system compared to hBCN - group. Conclusions:Env genes in hBCN + group and hBCN - group may have relatively unique amino acid usage patterns, and virus strains in hBCN + group are less adaptable to the host than those in hBCN - group.

Objective To explore the impact of broad antigen HLA-Bw4 on disease progression in HIV-1 infected subjects. Methods Three hundred and forty subjects chronically infected with HIV-1 and 69 HIV-1 negative subjects were recruited and HLA-B alleles were typed with sequence-based high resolution typing assay. HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD4+ T cell counts and viral loads were analyzed. Results Sixty-five HLA-B alleles were detected in HIV-1 positive subjects. Subjects with Bw4 (Bw4 homozygotes and Bw4Bw6 heterozygotes ) had higher CD4+ T cell counts ( P = 0. 004 ) and lower plasma viral load ( P = 0.003 ) than subjects without Bw4 ( Bw6 homozygotes). When compared with HIV-1 postive subjects with CD4+ T cell counts above 500 celis/μl, those with CD4+ T cell counts below 500 cells/μl were observed with decreased percentage of Bw4Bw6 heterozygote ( P =0.0002) and increased percentage of Bw6 homozygotes ( P < 0. 0001 ). There is no significant difference in CD4+ T cell counts between Bw4 homozygotes and Bw4Bw6 heterozygote, but lower viral loads were observed in Bw4Bw6 heterozygotes( P = 0. 037 ). Conclusion HLA-Bw4 can confer pretective effects on H1V-1 infected subjects by maintaining higher CD4+ T cell counts and lower viral load, the mechanism behind this effect need further exploration.

Objective An ELISA-based assay for detecting alloantibodies against FⅧ was established to estimate the incidence of alloantibodies against FⅧ in treated patients with hemophilia A. Methods One hundred and forty patients with hemophilia A and 80 healthy controls were enrolled. Among hemophilia A patients, 84, 34 and 22 patients were in severe, moderate and mild conditions respectively. All patients were treated with plasma-derived FⅧ concentrates before. The titer wells were coated with MoAb against FⅧ which was developed in our laboratory. Then human recombinant FⅧ concentrates were applied. After incubation in room temperature for 2 hours, diluted plasma samples and HRP-conjugated goat anti-human IgG were added successively. Finally Absorbance (A490) were measured and recorded. Inhibitor activity against FⅧ for all plasma samples was measured by a modified Nijmegen assay simultaneously. Results The results showed that alloantibodies against FⅧ were found in 40.0% (56/140) patients by ELISA. And the alloantibody incidences in the severe and non-severe patients were 47.6% (40/84) and 28.5% (16/56)respectively. There was statistical significance between these two categories (x2 = 5.079, P ＜ 0.05 ). The FⅧ inhibitor activity was detected in 24.3% (34/140) patients by modified Nijmegen assay. The inhibitor incidences in the severe and non-severe patients were 33.3% (28/84) and 10.7% (6/56) respectively.There was statistical significance (x2 = 9.349, P ＜ 0.05). Twenty-five patients were positive for FⅧ alloantibodies by ELISA but had no FⅧ inhibitor activity by the modified Nijmegen assay. The positive rates of FⅧ alloantibodies and inhibitor activity were 40.0% (56/140) and 24.3% (34/140) respectively,which had significant difference (x2 = 15.75, P ＜ 0.01 ) and strong positive correlation ( rn = 0.59, P ＜0.01 ). Meanwhile the results deduced from these two tests shared a high consistency rate ( Kappa = 0.55,P ＜0.01 ). Conclusions The detection rate for alloantibodies against F Ⅷ is enhanced by our newlydeveloped ELISA. Our results suggest that the occurrence of the alloantibodies against F Ⅷ in Chinese hemophilia A patients is not rare and the alloantibody incidence is preponderant in the patients with severe hemophilia A compared with non-severe hemophilia A patients.

Objective To analyze characteristics of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype B/C infectors. Methods 19 HIV-1 recombinant subtype B/C infectors infected within 1 year, 40 chronic infectors infected for more than 3 years were enrolled in this cohort study. Elispot assay was used to observe HIV-1 specific T lymphocyte responses in HIV-1 recombinant subtype B/C infectors. Results Nef-specific T lymphocyte responses of interferon-gamma secretion were identified in 15 Chinese HIV-1 recombinant subtype B/C infectors infected within 1 year. The specific T lymphocytes were mainly targeted at four peptides which span from Nef 83 to 135: EVA7081.1, EVAT081.5, EVA7081.6 and EVAT081.48. Responses were identified in 29(75. 2%) infectors with more than 3 years of infection and the specific T lymphocytes were mainly targeted at three peptides which span from Nef 63 to 101 : EVA7081.43, EVA7081.44, EVAT081.45, EVA7081.47, EVA7081.48 and EVA7081.49. The average magnitude of response in infectors with less than 1 year of infection was 284. 13 SFC/106 PBMC. The average magnitude of response in infectors with more than 3 years of infection was 152. 44 SFC/106 PBMC. There was a significant difference between the two groups (U = 91. 000, P = 0. 002). Conclusions HIV-1recombinant subtype B/C infectors at different stages of diseases (less than 1 year and more thank 3 years) can recognize central region of Nef. The magnitude of Nef-specific IFN-γ secretion T lymphocyte responses in this cohort gradually decrease with disease progression.

Objective To analyze the character of Pol-specific T lymphocyte responses and identify immunodominant region recognized in Chinese HIV-1 recombinant subtype B/C infectors at different stages of diseases. Methods Eleven Chinese HIV-1 recombinant subtype B/C infectors infected in 18 months, 25 which infected time more than 3 years and 10 HIV-1-seronegative healthy individuals were enrolled. HIV-1-specific T lymphocyte responses were analyzed by an IFN-γ ELISPOT assay against 249 overlapping peptides spanning HIV-1 Pol protein in the present study. Results Pol-specific T lymphocyte responses of IFN-γsecretion were identified in 8 (72.73%) out of 11 infectors infected in 18 months, the specific T lymphocytes are mainly targe-ted at six peptides which amino acid position from Pol 481 to 631 in reverse transcriptase region: Pol5581, Pol5582, Pol5587, Pol5609, Pol5610 and Pol5615. There was a negative correlation between the breadth of re-sponse and peripheral CD4+ T cell count (P=0.0212, r=-0.762) ; Responses were identified in 15 (60%) out of 25 chronic infectors, the specific T lymphocytes are mainly targeted at four peptides which amino acid po-sition from Pol 241 to 295: Pol5521, Pol5525, Pol5526, Pol5531 and another peptide: Pol5638 which amino acid position from Pol 708 to 722 in reverse transcriptase region. There was a positive correlation between the magnitude of Pol-specific IFN-γ secretion T lymphocyte responses and plasma viremia (P = 0.006 95 , r = 0.660) . None of the seronegative healthy individuals gave the positive responses. Conclusion Chinese HIV-1 recombinant subtype B/C infectors at different stages of diseases mainly recognized different re-gions of Pol.