BACKGROUND: Biological function of leukocyte-associated immunoglobulin like-receptor 1 (LAIR 1) has clearly researched in China and abroad, but the in vivo biological function of LAIR is poorly understood. OBJECTIVE: To establish LAIR-2 (CD306) eukaryotic expression vectors and to purify and identify the fusion protein. DESIGN, TIME AND SETTING: A single sample observation experiment was performed at the Fourth Military Medical University of Chinese PLA between June 2007 and June 2008.MATERIALS: plg/3c vector was offered by Oxford University, pcDNA3.1 vector was provided by Meyaard doctor. Chinese hamster ovary (CHO) cell lines were preserved by the Department of Immunology, Fourth Military Medical University of Chinese PLA.METHODS: Two eukaryotic expression vectors plg/3c-LAIR-2 and pcDNA3.1-LAIR-2 were constructed and were transfected into CHO cells. The binding activities of LAIR-2 fusion protein to LAIR-2 mAbs were identified by Western blot, immunocytochemistry and flow cytometry assay.MAIN OUTCOME MEASURES: The construction of stably transfected cell lines, and the purification and identification of fusion protein. The activity of LAIR 2 protein combined to corresponding monoclonal antibody.RESULTS: Eukaryotic expression vectors were constructed and trasnsfected into CHO cells successfully. Two cells lines CHO/LAIR-2-Fc and CHO/LAIR-2 that steadily expressed LAIR-2-Fc fusion protein and LAIR-2 protein were established. Western blot assay showed that LAIR-2 protein could bind specially to LAIR-2 mAb 1A7, 3H12 and 4A9. Immunocytochemistry and flow cytometry assay demonstrated that 3H12 and 4A9 could bind to LAIR-2 expressed in the transfected CHO cells. CONCLUSION: Two ceils lines CHO/LAIR-2-Fc and CHO/LAIR-2 were successfully constructed, which can transfected to CHO cells. The eukaryotic expressed LAIR-2 protein has good binding activity to LAIR-2 mAbs.

Objective To study the relationship of soluble LAIR (sCD305 and CD3060) expression in recipient serum with cytomegalovirus (CMV) pneumonitis after renal transplantation. Methods Nineteen serum specimens from recipients were divided into CMV pneumonitis group (n=10) and control group (n=9). Then the concentrations of sCD305 and CD3060 were quantitated with sandwich ELISA. The data were analyzed by using student t test. Results sCD305 was skewness distributed in both 2 groups, was 0.000-3.039 μg/L in CMV pneumonitis group and 0.000-8.375 μg/L in con-trol group. CD3060 was skewness distributed in CMV pneumonitis group and the concentration was 0.000-0.017μg/L. CD3060 was mormally distributed in control group and the concentration was 0.046±0.035 μg/L. There was significant difference of CD3060 (P=0.000) concentrations and no sig-nificant difference of sCD305(P=0.316) concentrations in 2 groups, respectively. Conclusions The concentration of CD3060 is low in CMV pneumonitis patients. The combination of CMV PP65 antigen detection and CD3060 detection is helpful for the early and precise diagnosis of CMV pneumonitis in renal transplantation patients.

BACKGROUNDTo investigate the relationship between interleukin-6 (IL-6) levels in serum and HBV replication and the role of IL-6 in liver damage in chronic hepatitis B patients.

METHODSDetected IL-6 levels and polymerized human serum albumin receptor (PHSA-R) in serum of different type of HBV markers positive patients and of different degree of chronic hepatitis B patients.

RESULTSThe results showed that the serum IL-6 levels were increased significantly in PHSA-R positive patients than in PHSA-R negative patients in chronic hepatitis B (P<0.01). The serum IL-6 levels were correlated with the levels of PHSA-R (r=0.694, P<0.01), and the degree of symptoms.

CONCLUSIONSThis study suggested that the IL-6 levels in serum of hepatitis B patients correlated HBV replication and the degree of liver damage, serum IL-6 levels may be used as a value indicating of HBV replication, the degree of symptoms and the effect of treatment.

Adult ; Aged ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; blood ; Humans ; Interleukin-6 ; blood ; Middle Aged ; Receptors, Albumin ; blood ; Virus Replication

Objective To investigate the role of soluble CD305(sCD305)in renal allograft rejection.Methods Concentration of serum sCD305 was detected on 20 healthy volunteers and 153 cases of recipients after kidney transplantation by using double monoclonal antibody sandwich enzyme linked immunosorbent assay.Results In the healthy volunteers and 98 recipients with normal renal funetion,sCD305 was detected at low levels of(4.3±2.3)μg/L and(6.3±3.7)μg/L.In 20 cases of acute rejection and 5 cases of graft loss,serum sCD305 levels were(36.3±14.7)μg/L and(28.8±9.4)μg/L,and significantly higher than those in the healthy volunteers and recipients with normal renal function.Meanwhile,in the 30 cases of chronic rejection and 6 cases under dialysis treatment,the levels of sCD205were(13.1±5.5)ttg/L and(11.2±4.6)μg/L and significantly higher than those in the healthy volunteers and recipients with normal renal function.Conclusions CD305 was presented at high level in the recipients with renal acute or chronic rejection,and it might be a potential marker for monitoring graft rejection after transplantation.

Objective To investigate the relationship between the soluble LAIR-1(sLAIR-1)in the serum from recipients after transplant and graft rejection.Methods Serum sLAIR-1 level was determined by double mAb sandwich enzyme linked immunosorbent assay on 23 cases of liver transplantation and 139 cases of kidney transplantation.Results In healthy volunteers and 98 recipients with normal graft function,sLAIR-1 was detected at low level [(4.3±2.3)μg/L and(6.3±3.7)μg/L],with the difference being not significant.In 6 cases of liver acute rejection,20 cases of kidney acute rejection and 5 cases of graft loss,serum sLAIR-1 levels were increased remarkably at high 1evels [(47.2±25.9)μg/L,(36.3±14.7)μg/L,and(28.8±9.4)μg/L respectively]as compared with the two groups of healthy volunteers and the recipients with normal graft function,even peaked at 117.3 μg/L in one case of severe liver rejection.Meanwhile,in 5 cases of liver chronic rejection,27 cases of kidney chronic rejection and 6 cases under dialysis treatment.the levels of sLAIR-1 were(16.1±6.4)μg/L,(13.1±5.5)μg/L and(11.2±4.6)μg/L respectively,significantly higher than those of the healthy volunteers and the recipients with normal graft function.Conclusion sLAIR_1 was detected at high level in the recipients suffered graft acute or chronic rejection and might be a promising monitor of rejection after transplantation.

Objective Based on detection of the soluble LAIR-1 (sLAIR-1) and sIL-2R in the bile from recipient after liver transplant, the role of sLAIR-1 and sIL-2R in graft acute rejection were analyzed. Methods Bile sLAIR-1 level and sIL-2R were determined by double mAb sandwich enzyme linked immunosorbent assay in 55 cases of liver transplantation. Results In 22 recipients with normal graft function, sLAIR-1 and sIL-2R were detected at low level in the bile. In the 29 cases of liver acute rejection (AR), significant increase of bile sIL-2R level was detected on the lst and 2nd d before final diagnosis. With the effective methylprednisolone pulse therapy, sIL-2R level was decreased significantly on the 3rd d. On the other hand, remarkable increase of bile sLAIR-1 was found on the lst,2nd and 3rd d before final diagnosis. After of methylprednisolone pulse therapy for 3 d, bile sLAIR-1resturned to the control level. Conclusion Both bile sIL-2R and sLAIR-1 are detected at high level in the recipients suffering from liver acute rejection. The level of bile sLAIR-1 changes dramatically and responsively according to liver acute rejection. Therefore, detecting these two markers synergistically may be a promising monitor for rejection after liver transplantation.

Objectives:To determine the expressions of decoy receptors (DcR1 and DcR2) of TRAIL in carcinoma of endometrium. Methods:The expressions of DcR1 and DcR2 in endometrium tissues from 13 carcinoma of endometrium and 7 normal endometrium were detected by immunohistochemical staining.Results: The expressions of DcR1 and DcR2 in carcinoma of endometrium were much lower than in normal endometrium. Conclusions:The decreasing of DcR1 and DcR2 in carcinoma of endometrium may be concerned with its pathogenesis, which may be related to the prevention of endometrium from carcinomatous change.

BACKGROUND:Cytotoxic T lymphocyte-associated antigen 4 is a newly discovered costimulatory molecule. It has been studied more in tumor and autoimmune diseases, less in the field of kidney transplantation.
OBJECTIVE:To explore the role of cytotoxic T lymphocyte-associated antigen 4 in acute rejection after renal transplantation.
METHODS:Fifty patients undergoing renal transplantation were divided into acute rejection group (20 cases) and stable graft function group (30 cases). Another 30 healthy persons served as control group. Blood samples were extracted from the peripheral blood. Cytotoxic T lymphocyte-associated antigen 4 was detected by enzyme linked immunosorbent assay and flow cytometry.
RESULTS AND CONCLUSION:The expression of cytotoxic T lymphocyte-associated antigen 4 in the serum showed significant differences in the acute rejection group, stable graft function group and healthy control group (F=70.008 1, P=0.000 0), but showed no difference in peripheral blood lymphocytes of three groups (F=1.865 6, P=0.161 7). Compared with the healthy control group, the expression levels of cytotoxic T lymphocyte-associated antigen 4 in peripheral blood lymphocytes of acute rejection group and stable graft function group were significantly decreased (P=0.000 0). In addition, the acute rejection group had a lower cytotoxic T lymphocyte-associated antigen 4 expression than the stable graft function group (P=0.000 0). In renal transplant rejection, the expression of cytotoxic T lymphocyte-associated antigen 4 in serum was reduced, showing some correlation with acute rejection after renal transplnatation. Cytotoxic T lymphocyte-associated antigen 4 might be involved in the rejection.

Objective To evaluate the clinical application of female orthotopic ileal neobladder.Methods Modified radical cysteetomy plus orthotopic ileal neobladder was performed on 19 female pa-tients with bladder cancer from June 1999 to January 2008.The mean age of the patients was 52(45-66) years,mean course of disease was 4.4 months (16 days-1.9 years).Of all the patients,there were 10 cases with grade 1,7 cases with grade 2 and 2 cases grade 3.According to the UICC stage system,5 patients were T1 stage,12 T2 and 2 T3a.All the patients received modified radical cystecto-my without resection of uterus and anterior vagina,meanwhile the nerves around urethra were protec-ted.0.8-1.2 cm proximal end of the urethra was excised and 30 cm distal ileum was used for the re-construction of the neobladder.Results Sixteen cases were followed up for 6-102 months,mean 71 months.Fifteen patients survived without disease recurrence,1 patient died of myocardial infarc-tion 17 months postoperation.The daytime and night continent rate was 100$,93% at 9 months postoperative.The average voiding volume of the 15 patients was 519.0 ml.The average residual vol-ume was 29.2 ml,and Qmax was 18.6 ml/s.The average filling and voiding pressure was 16.7 cm H2O and 53.0 cm H2O.Intravenous urography showed slight hydronephrosis in 1 case.Conclusion Female orthotopic ileal neobladder could be a good choice because of the continence,fewer complica-tions,lower pressure and enough bladder capacity.

Objective:To detect the expression of ALG3 in human bladder cancer and analyze its clinical significance.Methods:Bioinformatics was conducted to analyze the mRNA level of ALG3 in bladder cancer tissues and normal tissues, and to analyze the relationship between its expression and the survival rate of bladder cancer patients. The clinical and pathological data of 87 patients with bladder cancer who underwent surgical treatment were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of ALG3 protein in bladder cancer tissues and healthy tissues adjacent to the cancer, and to analyze its relationship with clinicopathological characteristics of bladder cancer patients.Results:The results of bioinformatics analysis showed that ALG3 mRNA was highly expressed in bladder cancer tissues, and was significantly correlated with the overall survival rate and disease-free survival rate of patients. Immunohistochemical results showed that ALG3 was mainly localized in the cytoplasm and was highly expressed in bladder cancer tissues, but low or no expression in healthy tissues adjacent to the cancer. The high expression of ALG3 in bladder cancer tumor tissue was related to tumor stage and tumor recurrence (all P<0.05), but not related to age, gender, tumor grade and lymph node metastasis (all P>0.05). Conclusions:ALG3 plays an important role in the occurrence and development of bladder cancer, and it is expected to become an important therapeutic target and prognostic marker for bladder cancer.