Background Interleukin-17 (IL-17)is a potent pro-inflammatory cytokine and plays a pathogenic role in autoimmune disease.It was confirmed that IL-17 is implicated in allograft rejection of many transplanted organs.Recent studies have foensed on the effect of IL-17 antagonists on allograft rejection.Objective This study aimed to investigate the inhibitory effect of anti-mouse IL-17 monoclonal antibody (mAb) on corneal allograft rejection.Methods Twenty-five 8 to 10-week-old C57BL/6 mice and 50 BALB/c mice were collected.Donor cornea grafts with 2 mm diameter from 25 C57BL/6 mice was transplanted to 50 eye of BALB/c mice to establish a model of corneal transplantation.The recipients were randomized into 2 groups,and neutralizing mouse IL-17antibody or isotype control antibody was intraperitoneally injected immediately after transplantation for experimental treatment,respectively.Allografts were scored clinically at appropriate time points after treatment based on Plskova criteria,and ≥5 was confirmed as rejection.Infiltrating cells in corneal graft were detected qualitatively and quantitatively by immunohistochemistry and reverse transcription-PCR separately.The cytokine levels of T helper type 1 (Th1),Th2,and Th17 in recipients' spleen wer(c) analyzcd by ELISA.The use of the animals followed the Statement of ARVO.Results Compared with the isotype control antibody group,the survival of grafts was improved in the IL-17mAb group(P＜0.05).The levels of neutrophile granulocyte mRNA,CD4+ and CD8+ T lymphotes mRNA were 2.22±0.10,1.64±0.04 and 1.32±0.10 in the IL-17 mAb group,showing a significant decline in comparison with those of the isotype control antibody group(3.61 ±0.08,2.69±0.06 and 2.17±0.04) (P=0.000,0.000,0.000).Interferon-γ(IFN-γ),IL-12 p40 and IL-17 concentrations in recipients ' splenocytes were (529.80 ± 13.83) ng/L,(539.58 ±10.74) ng/L and(173.70±8.11)ng/L in the IL-17 mAb group,and thosc in the isotype control antibody group were (741.48± 10.51) ng/L,(1156.90 ± 69.93) ng/L and (366.13± 7.93) ng/L,with significant differences between them (P=0.000,0.001,0.000).Conclusions Neutralization IL-17 bioactivity inhibits mouse corneal allograft rejection to a certain extent.

OBJECTIVEMcAbs against rabies nucleocapsid were used to detect rabies street viruses in animal brain specimens with indirect immunofluorescent assay to evaluate the sensitivity and specificity of this assay.

METHODS62 specimen from rabid animal brains including genotype 1 to 7 and 271 specimens from different normal animal brains collected in Pasteur Institute in 2003 were tested and compared, using indirect immunofluorescent assay. All these specimens were identified and compared using rapid rabies enzyme immunodiagnosis, fluorescent antibody test and rabies virus isolation assay in neuroblastoma cell culture which were all provided by Pasteur Institute.

RESULTSBoth sensitivity and the specificity of the indirect immunofluorescent assay were 100%.

CONCLUSIONThe results showed a positive of rabies virus detection with these methods.

Animals ; Antibodies, Monoclonal ; immunology ; Antibodies, Viral ; immunology ; Brain ; virology ; Dogs ; Fluorescent Antibody Technique ; methods ; Genotype ; Nucleocapsid ; immunology ; Rabies virus ; isolation & purification ; Sensitivity and Specificity

This study was aimed to explore clinical significance of Livin mRNA and protein expressions in non-Hodgkin's lymphoma (NHL). The immunohistochemistry was used to determine the expression of Livin protein in lymph nodes of 30 patients with NHL and 11 patients with reactive hyperplasia of lymph node. The real-time PCR was performed to detect the expression levels of Livin mRNA in 20 patients with NHL, 10 patients with reactive hyperplasia of lymph node and 4 normal person lymph nodes. The correlation of Livin mRNA and protein expressions with NHL clinical features were analyzed. The results showed that the expression of Livin mRNA was statistically higher in NHL samples than in normal lymph nodes and reactive hyperplastic lymph nodes (12.4 vs 0.34, 12.4 vs 0.61, median) (p<0.05), while there was no statistical difference between normal and reactive hyperplastic lymph nodes (p>0.05). Livin protein expression was exhibited to be positive in 16 of 30 cases of NHL with a positive rate of 53.3% and only 1 in reactive hyperplastic lymph nodes with a positive rate of 9.1%. In addition, Livin protein almost appeared in the cytoplasm of cells, seldom in nucleus. The expressions of both Livin mRNA and protein were positively correlated with clinical stages of NHL (p=0.023; p=0.009), B symptoms (p=0.015; p=0.026), blood beta2-microglobulin (beta2-MG, p=0.031; p=0.012) and the serum level of lactate dehydrogenase (LDH, p=0.037; p=0.007), but the expressions of Livin mRNA and protein had no significant correlation with age, sex and typing. It is concluded that the Livin mRNA and protein highly express in NHL patients and correlate with many clinical features, such as stage of NHL, B symptom, beta2-MG and LDH, therefore, the Livin may play a role in the prognosis of NHL patients. The further study on inhibitory effect of Livin expression will promote the illustration of NHL pathogenesis and contribute to the treatment and prognosis of NHL.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adult ; Aged ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Lymph Nodes ; pathology ; Lymphoma, Non-Hodgkin ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Neoplasm Staging ; RNA, Messenger ; genetics

This study was aimed to investigate the expression and clinicopathologic significance of Gli1 and Gli2, 2 factors of Hedgehog(Hh) signaling pathway, in non-Hodgkin's lymphoma (NHL). Gli1 and Gli2 mRNA and protein in 18 cases of NHL and 10 cases of reactive lymphadenitis were amplified and identified by real-time PCR, and were assayed by immunohistochemical staining respectively. The results showed that (1) Gli1 and Gli2 mRNA in NHL group (RQ 2.05, 2.31) were expressed higher than that in reactive lymphadenitis group (RQ 0.82, 0.89). Gli1 mRNA activated level was positively related with Gli2 (r = 0.63, p < 0.01). In addition, Gli2 also positively correlated to clinical stages of NHL (p = 0.03), but the expressions of Gli1 and Gli2 mRNA had no significant correlation to B symptoms, blood β(2)-microglobulin, age and sex. (2) The positive expression rate of Gli1 and Gli2 protein in NHL group were 80% and 68% respectively, which were extremely higher than that in reactive lymphadenitis group. Gli1 protein level was positively related with Gli2 (r = 0.62, p < 0.05). Both Gli1 and Gli2 protein expression positively correlated to clinical staging of NHL (p = 0.05, p = 0.01). It is concluded that the Gli1 and Gli2 of Hh signaling pathway have been found to higher express in patients with NHL, and have significance for clinical staging and predicting prognosis of NHL. To further investigate the role of Hh signaling pathway in NHL will contribute to elucidate the occurrence and development of NHL, and provide a favorable method for therapy of NHL.
Adult ; Aged ; Female ; Hedgehog Proteins ; metabolism ; Humans ; Kruppel-Like Transcription Factors ; metabolism ; Lymphoma, Non-Hodgkin ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Nuclear Proteins ; metabolism ; Signal Transduction ; Transcription Factors ; metabolism ; Zinc Finger Protein GLI1 ; Zinc Finger Protein Gli2

OBJECTIVETo investigate the expression of human epididymal secretary protein 2 isoform human epididymal protein 2beta1(HE2beta1) in the testis and epididymis of adolescent male rats along with its significance.

METHODSImmunohistochemical staining was used to detect the expression and localization of HE2beta1 in the testis and epididymis of 15 adolescent SD rats.

RESULTSHE2beta1 immunoreactive staining was detected in the testis and epididymis. In the epithelia of the epididymal duct, HE2beta1 expressed mainly in the supranuclear region of the principle cells and the basement membrane of some epithelial cells; there were no immunostaining in the n clear cells, halo cells and basal cells. The immunopositive reaction was detected, weak in the distal caput, strong in the proximal, middle corpus and the cauda, but negative in the initial segment. Immunopositive results of HE2beta1 were also observed in some of the nuclei of spermatogonia and Sertoli cells with negatively-stained cytoplasm.

CONCLUSIONImmunohistochemical staining is a fairly sensitive method for detecting HE2beta1 expression. The localization and expression level of HE2beta1 in the genital duct of adolescent male rats exhibited a region- and cell-specific expression pattern, which suggests that HE2beta1 may play an important role in spermatogenesis, maturation and epididymal epithelial innate defense mechanisms.

Animals ; Antigens, Surface ; biosynthesis ; Epididymis ; metabolism ; Glycopeptides ; biosynthesis ; Humans ; Immunohistochemistry ; Leydig Cells ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism

Objective Ambulatory arterial stiffness index (AASI) has been recently proposed to reflect the dynamic relation between diastolic and systolic blood pressure throughout the whole day. The aim of our study was to investigate the change in AASI with advancing age and the correlation with 24 hours pulse pressure (24 h PP) in healthy individuals. Methods 246 healthy subjects [mean age(59. 7 ±14.6) years, women 38.6% ] underwent 24 hours ambulatory blood pressure monitoring ( ABPM) in normal life style. The blood pressure recordings, heart rate (HR) , mean arterial pressure (MAP) and pulse pressure (PP) were analyzed simultaneously by computer for every 30 minutes during 6:00 am-22:00 pm and every 60 minutes during 22:00 ptn-6:00 am. Using all the blood pressure recordings, we plotted diastolic against systolic blood pressure from each individuals and calculated the regression slope. AASI was derived from 1 minus this regression slope. Results In 246 healthy individuals, AASI increased with age. Among the healthy individuals, the 95th percentile of AASI was 0. 56, the upper boundary of the 95% prediction interval of AASI in relation to age were 0.49 at 20 - 39 years ,0.59 at 40 - 59 years ,0.69 at 60 - 79 years, 0.79 at&ge;80 years. The correlation coefficient between AASI and 24 h PP was 0.497(P <0. 01 ). AASI linearly increased with age in healthy individuals, whereas the relation between pulse pressure and age was curvilinear. Conclusions AASI as a index reflecting blood pressure relationship, manifested the corresponding change with advancing age. The correlation between AASI and traditional index 24 h PP indicated AASI as a new measure of arterial stiffness.

Background::After neoadjuvant chemotherapy (NAC), non-pathological complete response of breast cancer patients can benefit from tailored adjuvant chemotherapy. However, it is difficult to select patients with poorer prognosis for additional adjuvant chemotherapy to maximize the benefits. Our study aimed to explore whether the subtypes of tumor-infiltrating lymphocytes (TILs) in residual tumors (RT) is related to the prognosis of triple-negative breast cancer (TNBC) after NAC.Methods::Data from patients with primary TNBC consecutively diagnosed at the Breast Disease Center of Peking University First Hospital from 2008 to 2014 were retrieved, and the cases with RT in the breast after NAC were enrolled. TILs subtypes in RT were observed by double-staining immunohistochemistry, and counted with the median TILs value per square millimeter as the cut-off to define high versus low TILs density in each subtype. The relationships between the TIL density of each subgroup and the clinicopathological characteristics of the RT after NAC patients were analyzed by Fisher exact test. Disease-free survival (DFS) and overall survival (OS) were analyzed by the Kaplan-Meier method and log-rank statistics.Results::A total of 37 eligible patients were included in this study, and the median follow-up period was 50 months (range 17–106 months). There was no significant correlation between the infiltrate density of CD4 +, CD8 +, CD20 +, and CD68 + lymphocytes and clinic-pathological characteristics. Significantly better prognosis was observed in patients with high CD4 +-TILs (DFS: P = 0.005, OS: P = 0.021) and high CD8 +-TILs (DFS: P = 0.018) and low CD20 +-TILs (OS: P = 0.042). Further analysis showed that patients with CD4 +/CD20 + ratio greater than 1 (DFS: P = 0.001, OS: P = 0.002) or CD8 +/CD20 + ratio greater than 1 (DFS: P = 0.009, OS: P = 0.022) had a better prognosis. Conclusions::Subtypes of TILs in RT is a potential predictive biomarker of survival in TNBC patients after NAC.

OBJECTIVETo assess the efficiency of eluting stent coated with arsenic trioxide (As(2)O(3)) suspended in poly-L-lactic acid (PLLA) to prevent in-stent restenosis in rabbits.

METHODSForty-five male New Zealand white rabbits were assigned to three groups (n = 15 for each group) at random: uncoated stents, stents coated with PLLA or stents coated with As(2)O(3) in PLLA. Animals were euthanized 28 days after stent implantation into the iliac arteries of rabbits. Neointimal thicknesses and apoptosis of vascular smooth muscle cell (VSMC) were measured. Stents coated with As(2)O(3) in PLLA were implanted in another 48 male New Zealand white rabbits, As(2)O(3) concentrations in serum and arterial tissue at implantation site were measured at 2 h and 1, 3, 7, 14, 28 days after As(2)O(3) eluting stent implantation (n = 8 for each time point).

RESULTSNeointimal hyperplasia was significantly reduced 51% and 31% and apoptosis significantly increased (21.0 +/- 3.3; 6.2 +/- 1.9(*); 5.3 +/- 2.1(*), (*)P < 0.01 vs. As(2)O(3) eluting stent) with As(2)O(3) eluting stent, versus PLLA-coated stents and uncoated stents. As(2)O(3) concentrations in arterial tissue at implantation site were 18.6 +/- 9.1 (ng/mg) at 1 day and 0.3 +/- 0.1 (ng/mg) at 28 days after stent implantation.

CONCLUSIONSAs(2)O(3) coated stents released As(2)O(3) to local tissue for at least 28 days, suppressed neointimal hyperplasia in rabbit iliac arteries and increased local VSMC apoptosis might be one of the mechanisms for inhibiting restenosis by As(2)O(3) coated stents.

Animals ; Apoptosis ; drug effects ; Arsenicals ; administration & dosage ; Coronary Restenosis ; prevention & control ; Drug-Eluting Stents ; Iliac Artery ; Male ; Muscle, Smooth, Vascular ; cytology ; Oxides ; administration & dosage ; Rabbits ; Random Allocation

OBJECTIVETo explore the Notch1 mRNA and protein expression in human breast cancers and normal mammary tissues, and their relationship with the clinical indicators of breast cancers were analyzed.

METHODSNotch1 gene of human breast invasive ductal carcinoma (IDC) and normal mammary gland tissues were amplified by RT-PCR, and the expression of Notch1 protein was detected by immunohistochemical Streptavidin-Biotin Complex (SP) stain in 60 IDC, 30 ductal carcinoma in situ (DCIS) and 60 normal mammary tissues.

RESULTSNotch1 gene of human IDC and normal mammary tissues both could express in a transcription level; the positive rates of Notch1 protein expression in normal mammary tissues and DCIS were 55% and 70%. Respectively, which did not differ statistically (P > 0.05), while the positive rate in IDC was 90%, significantly higher than that of the normal mammary tissues and DCIS (P < 0.05). The high expression of Notch1 protein in IDC correlate significantly with lymph node metastasis, pathological grades and TNM stages.

CONCLUSIONSNotch1 protein was over expressed in breast IDC. A high Notch1 protein expression is considered associating with the evolution and malignant transformation of the breast tumor. The expression of Notch1 gene maybe impact the effect of on the progression of breast cancers.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Mammary Glands, Human ; metabolism ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Receptor, Notch1 ; genetics ; metabolism

OBJECTIVETo study the impact of an agonist anti-CD(40) monoclonal antibody 5C11 on the induction and biological characteristics of leukemic dendritic cells.

METHODSCombinations of 5C11 and different cytokines were used to induce differentiation of leukemic blasts into dendritic cells. Morphology was observed by light microscopy. Surface antigens of the induced cells were analyzed by fluorescence-activated cell sorting (FACS), the yields of dendritic cell by cell counting, the levels of IL-6 and IL-12 by ELISA, T cell proliferating activity by allo-mixed lymphocyte reaction (MLR) in vitro. Allogeneic T cells were stimulated with leukemic dendritic cells and T-cell cytotoxicity was measured by MTT assay.

RESULTSWhen cultured with combinations of 5C11 and different cytokines, the leukemic cells isolated from the patients could differentiate into dendritic cells. The morphology showed typical features of dendritic cells, which expressed high levels of CD(40), CD(80) and CD(86). In comparison with the original leukemia cells, the leukemic dendritic cells secreted less IL-6 but more IL-12 (P < 0.05). The leukemic dendritic cells were potent to stimulate the proliferation of allogeneic T cells, and the latter was able to lyse the original leukemia cells.

CONCLUSIONLeukemic blasts could be induced to differentiate into functional dendritic cells. It may be of great value in the adoptive immunologic therapy of leukemia.

Antibodies, Monoclonal ; immunology ; CD40 Antigens ; physiology ; Cell Differentiation ; Dendritic Cells ; immunology ; Humans ; Immunophenotyping ; Immunotherapy ; Interleukin-12 ; biosynthesis ; Interleukin-6 ; biosynthesis ; Leukemia ; immunology ; pathology ; therapy