Changes in serum C3d and C5b-9 levels in elderly patients with idiopathic membranous nephropathy and their clinical significance / 中华老年医学杂志

Objective:To investigate changes in serum C3d and C5b-9 levels in elderly patients with idiopathic membranous nephropathy(IMN)and their correlations with prognosis.Methods:Two hundred thirty-one elderly patients with IMN and 96 non-elderly patients with IMN confirmed by kidney biopsy at the First Affiliated Hospital of Zhengzhou University from January 2015 to May 2017 were enrolled.During the same period, 118 healthy individuals receiving health checkups were included as controls.Patients were divided into the low C3d group( n=112)and the high C3d group( n=113)according to the median level of serum C3d.Serum C3d and C5b-9 levels were measured by enzyme-linked immunosorbent assays. Results:Serum C3d and C5b-9 levels in elderly IMN patients were 0.23(0.15, 0.45)mg/L and 0.28(0.20, 1.23)mg/L, respectively, which were higher than those in healthy controls[0.18(0.13, 0.22)mg/L, 0.22(0.16, 0.26)mg/L, respectively]( Z=-4.261 and -6.213, P<0.001). Serum C3d levels in elderly and non-elderly IMN patients were correlated negatively with the estimated glomerular filtration rate( r=-0.155 and -0.426, P=0.019 and 0.000), but positively with serum creatinine, anti-phospholipase A2 receptor(PLA2R)antibody levels and 24 h urinary protein( r=0.184, 0.326, 0.407, 0.321 and 0.145, P=0.005, 0.001, 0.000, 0.001 and 0.027). Kaplan-Meier survival analysis showed that the cumulative renal survival rate in elderly IMN patients was lower in the high C3d group than in the low C3d group(47.8% vs.70.8%, Log Rank χ2=7.399, P=0.007). Multivariate Cox regression analysis showed that high C3d levels were an independent risk factor for poor renal outcomes in elderly IMN patients( HR=2.288, 95% CI: 1.082-4.839, P=0.030). Conclusions:High serum C3d levels are associated with increases in urinary protein excretion and anti-PLA2R antibody levels, renal function decline, and poor renal outcomes in elderly IMN patients.

Screening and identification of human anti-C3d single chain antibodies / 国际药学研究杂志

Objective: To screen human single chain antibodies against human C3d from phage-displayed single-chain variable fragment(scFv)library, and analyze their binding activities. Methods: The phage display library was exposed to 3-round selections in immunotubes coated with recombinant C3d at a decreasing concentration range and progressively stringent washing conditions. The positive clones were identified by ELISA, followed with sequencing to determine the specific genes which were then cloned intoex-pression vectors. The single chain antibodies were expressed in the FreeStyleTM 293-F system and harvested by affinity purification. The binding activities with recombinant C3d were determined by using Bio-Layer Interferometry technology. Results: These experiments resulted in three novel single chain antibodies(that is, A1, A3 and B6)against C3d with high affinity ranging from 22.7 to 171 pmol/ L. Conclusion: The high affinity human single chain antibodies against human C3d were obtained.

Comparison of Iso-C 3D navigation system versus X-ray fluoroscopy in minimally invasive surgery for Kümmell disease / 中华创伤骨科杂志

Objective To compare the Iso-C 3D navigation system versus the traditional X-ray fluoroscopy in minimally invasive surgery for Kümmell disease.Methods Included for the present retrospective study were 42 patients with Kümmell disease who had been treated by percutaneous kyphoplasty (PKP) or percutaneous vertebroplasty (PVP) at Department of Spine Surgery,Honghui Hospital from October 2016 to June 2018.They were 11 men and 31 women,aged from 43 to 72 years (average,57.4 years).The operation was aided by Iso-C 3D navigational system in 21 patients (observation group) and by X-ray fluoroscopy in the other 21 patients (control group).The 2 groups were compared before operation,one day and 6 months after operation in terms of visual analogue scale (VAS),Oswestry disability index (ODI),cobb angle and anterior vertebral height.Results The 2 groups were comparable because there were no significant differences between them in the preoperative general data (P ＞ 0.05).All the patients were followed up for an average of 9.6 months (from 6.0 to 18.5 months).At one day after operation for the observation andcontrol groups,the VAS scores (2.2 ±0.7 and 3.9 ± 1.5),ODI scores (15.6 ±2.8 and 26.2 ±6.9),cob angles (19.5° ± 1.7° and 20.8° ± 0.8°) and anterior vertebral heights (20.7 ± 1.4 and 18.7 ± 1.5 mm) were all significantly improved compared with those before operation (P ＜ 0.05).At 6 months after operation for the observation and control groups,the VAS scores (1.6±0.6 and 2.7±1.0),ODI (14.2±3.1 and 21.5 ±4.2),cob angles (18.6°± 1.8° and 19.0°± 1.7°) and anterior vertebral heights (19.9 ± 1.8 and 17.8 ± 1.6 mm) were not significantly different from those at one day after operation (P ＞ 0.05).The VAS,ODI,cobb angle and anterior vertebral height at one day after operation for the observation group were all significantly better than those for the control group (P ＜ 0.05).The VAS,ODI and anterior vertebral height at 6 months after operation for the observation group were significantly better than for the control group (P ＜ 0.05),but there was no significant difference between the 2 groups in the cobb angle (P ＞ 0.05).No complications were observed in the observation group but 6 cases in control group reported cement leakage.Conclusion In the PKP/PVP treatment of Kümmell disease,Iso-C 3D navigational system may be superior to traditional X-ray fluoroscopy in that it can lead to better recovery of anterior vertebral height,kyphosis correction,pain relief and quality of life.

Value of cell-bound complement activation products in the diagnosis of systemic lupus erythematosus / 临床检验杂志

Objective@#To investigate the values of T lymphocyte-bound complement activation products such as T-C3d and T-C4d, B lymphocyte-bound complement activation products such as B-C3d and B-C4d and erythrocyte-bound complement activation products such as E-C3d and E-C4d in the diagnosis of systemic lupus erythematosus (SLE). @*Methods@#Peripheral blood samples from 68 SLE patients, 70 patients with non-SLE autoimmune diseases and 68 healthy controls were collected randomly, and the expression levels of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d in these samples were detected by flow cytometry. Meanwhile, antinuclear antibodies (ANA), anti-double stranded DNA antibodies (anti-dsDNA), peripheral blood cell count and other markers were also detected. The differences of cell-bound complement activation products in three groups were analyzed with the area under the receiver operating characteristic curve (AUC), nonparametric test, sensitivity and specificity. @*Results@#The specific median fluorescence intensity (SMFI) of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d in SLE patients were significantly higher than those in the patients with non-SLE autoimmune diseases and healthy controls (all P＜0.05). The SMFI (median \[P 25, P 75\]) of T-C4d, B-C4d and E-C4d in SLE patients were 3.8(1.2, 13.1), 22.1(6.2, 67.9) and 19.6(1.8, 52.4), respectively. The SMFI of T-C4d, B-C4d and E-C4d in SLE patients with reduced red blood cells and/or lymphocytes were significantly higher than that with normal red blood cell and lymphocyte count (all P＜0.05). The AUCs of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d were 0.711, 0.763, 0.663, 0.631, 0.611 and 0.615, respectively (all P＜0.05). The sensitivity of the combination of T-C4d with B-C4d (73.5%) in the diagnosis of SLE was superior to that of anti-dsDNA (36.8%). @*Conclusion@#The cell-bound complement activation products (CB-CAPs) are specifically expressed in SLE patients, and their combination detection is helpful for the diagnosis of SLE.

Expression of C3d in normal human liver tissues with non-immunologic osmostic mechanism / 中华病理学杂志

Objective@#To observe the deposition of complement C3d at different development stages in human normal organs and tissues, and investigate the significance of its deposition.@*Methods@#Using immunohistochemical methods, the deposition of C3d was detected at different development stages of 60 normal human organs and tissue specimens and double staining was performed in some specimens. Ninty-five cases of other organs or tissues were collected as control group.@*Results@#In 50 of 60 livers, it was observed the deposition of C3d in Glisson′s capsule and periportal sheath, with irregular linear network-like disposition surrounding the portal sheath. In different age groups, the expression of C3d was more beyond the 20 year-old group than 3 to 20 year-old group. There wasn′t any expression of C3d under 3-year-old group. Under the immuning electron micrograph, C3d depositing at the Glisson′s capsule was observed, without immuning compounding. Thirty in 40 spleens, deposition of C3d in capsules, arteries of lymphatic sheath, follicles in the spleen was observed.@*Conclusions@#The deposition of C3d in Glisson′s capsule, splenic trabeculae, fibrous sheath, endarterium of liver and spleen arterioles, within normal human tissues from patients elder than 3 years, are osmosis/immunogenic deposition. The deposition of C3d is a normal physiological phenomenon, and treatment of the deposition of C3d should be avoided, as it is an immune complex or immuning reaction phenomenon.

Usefulness of Additional LISS/Coombs Card Test with Enzyme-Treated Red Cells in Detecting Anti-Kidd Antibodies Not Detectable by NaCl/Enzyme Card Test Alone / 대한수혈학회지

BACKGROUND: Detection of anti-Kidd antibody is important because of its clinical significance. If detection is difficult due to weak serological reactivity or dosage effect, use of an enzyme method could be helpful. However, despite use of an enzyme method, we still observed weak reactivity of anti-Kidd antibody. METHODS: All identified anti-Kidd antibody cases from Jan 2012 to Aug 2015 in Asan Medical Center were reviewed. Antibody identification test was performed using the column agglutination technique using Bio-Rad ID-DiaPanel with LISS/Coombs card, Bio-Rad ID-DiaPanel-P with NaCl/Enzyme card, and ID-DiaPanel-P with LISS/Coombs card. The test results were compared. RESULTS: Sixty cases of anti-JK(a) or anti-Jk(b) were detected and tested by enzyme method. Among them, 34 (56.6%) cases showed strengthened reactivity using the ID-DiaPanel-P with NaCl/Enzyme card method. However, 26 (43.4%) cases showed weakened reactivity. Of these, 13 cases that could be tested by an additional method using ID-DiaPanel-P with LISS/Coombs card containing anti-IgG and anti-C3d showed successfully strengthened reactivity. CONCLUSION: The reactivity of anti-Kidd antibodies that was not strengthened using ID-DiaPanel-P with NaCl/Enzyme card method could be successfully strengthened by use of the ID-DiaPanel-P with LISS/Coombs card.

Serum C3d levels in tropical pulmonary eosinophilia.

Background & objectives: Results of earlier studies to evaluate the possible role of complement system in tropical pulmonary eosinophilia (TPE) using classical methods like serum haemolyte component CH50, C3 and C4 levels were inconclusive. In this study we determined levels of serum C3d which is a catabolic fragment of C3, to find out any direct evidence of activation of the complement system in TPE. Methods: The study population consisted of 3 groups. Group A consisted of 37 patients with well characterized TPE. In group B, 26 patients with pulmonary eosinophilia had similar respiratory and haemotological features as in Group A but had associated worm infestation in stool. The control group consisted of 39 healthy volunteers. Serum C3d levels were determined by sandwich ELISA technique. Results: The serum C3d levels in TPE patients were not significantly different from those of the patients of group B or the normal controls. Interpretation & conclusions: Absence of significant change in serum C3d goes against the possibility of complement activation in TPE. Results of our study suggest that complement system is unlikely to play a pivotal role in pathogenesis of TPE.

Molecular adjuvant C3d can improve humoral immunity of hCG? DNA immunization / 中国免疫学杂志

Objective:In the past study, a plasmid of pcDNA3 hCG? C3d3 had been constructed. It was suggested that the fusing protein be expressed by the pcDNA3 hCG? C3d3 plasmid both in the transient expression system in COS 7 cells, and the stable expression system in CHO cells. In the present research, it would be testified that the C3d molecular adjuvant could improve the immunogenicity of the hCG? DNA immunization or not. Methods:BALB/C mice of 6 weeks old were immunized intramuscularly two times at interval of 3 weeks by the plasmid pcDNA3, pcDNA3 hCG?, pcDNA3 hCG? C3d3 at dosage of 5, 10, 20 pmol, respectively. The anti hCG? antibody titers were determined by indirect ELISA in 6 weeks of last immunization. Results:The result showed that the C3d molecular adjuvant could increase significantly the titer of anti hCG? antibody in a dose dependent manner after DNA immunization.Conclusion:The C3d molecular adjuvant does improve the humoral immunity of the hCG? DNA immunization, which would be helpful for technical progress and clinical trial of the contraceptive vaccine. [

Expression of membrane complement regulatory protein on the surface of CD16~+ monocyte/macrophage and level of C3d in the blood of patients with glomerulonephritis / 中国免疫学杂志

Objective:To investigate the expression of membrane cofacto protein(MCP)?decay accelerating factor(DAF) and homologous restricition factor 20(HRF 20) on the surface of CD16 + monocyte/macrophage(Mo/M?) in patients with glomerulonephritis(GN).Methods:Tested the expression levels of MCP?DAF and HRF 20 on the surface of CD16 +Mo/M? by Flow Cytometry and the levels of serum C3d and C3d immunologic complex(C3d IC) by ELISA in 136 patients with GN,divided into mini change(MC)?glomerulosclerosis(GS)?membranous nephropathy(MN) and proliferative glomerulonephritis(PGN).Results:The expressions of MCP?DAF and HRF 20 on the surface of CD16 +Mo/M? and the serum levels of C3d in the patients with GS?MN or PGN were significantly higher than those in controls(P