Animals ; Arrestins ; metabolism ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; beta-Arrestins

Objective:To explore the characteristics of human melanoma-specific antigen peptides by HLA-A2 restricted tumor-infiltrating lymphocytes.Methods:The HLA-A2 protein and polypeptides molecules were purified from the three tumor cell lines(624-Mel, Chap-Mel and JY) by immunoaffinity chromatography, after the peptides bound to HLA-A2 protein solution were acidified with acetic acid and boiled by high temperature, and centrifuged through an Ultra-CL filter, then the peptides extracts were fractionated by revered phase high pressure liquid chromatography(RP-HPLC). Individual fractions were assessed for their ability to reconstitute melanoma-specific epitopes by adding to the HLA-A2 Ag-procceing mutant cell, T2. The biological feature of one of three active peptides from RT-HPLC samples was performed by mass spectrometric analysis. The synthetic peptides identical to active peptide sequences were determined in the reconstitute test.Results:Three prominent peaks(P19, P25 and P31) of the fraction from 624-Mel were observed in the reconstitute test, TIL killing rate was 67% for (P31) peptide fraction. The mass spectrometric analysis of one of active peptides (P31) showed that at mass-to-charge ratio(m/z) 948 has been usually nine residues. The sequence is H+ Ala Lue Trp Lue Phe Phe Gly Val Lue OH-. The peptide synthesized comprising epitopes were verified.Conclusion:These results showed the peptides derived from active fractions were related to human melanoma-specific tumor antigen peptides recognized by HLA-A2-restriced TIL. These peptides could develop novel peptide-based an anti-tumor vaccine for immunotherapy of CTL.

BACKGROUND:Poor implant anchorage in osteoporotic bone impacts its stability and requires the new solutions for the treatment. The augmentation technique with bone cements or bone substitutes is one strategy for the solutions. OBJECTIVE:To evaluate the transient stability of pedicle screw augmented using calcium sulfate cement in osteoporotic vertebral body. METHODS:Fresh calf lumbar vertebrae were selected to measure bone density, and then classified into four groups:the group by pedicle screw in normal vertebral body;the group by pedicle screw augmented using calcium sulfate cement in normal vertebral body;the group by pedicle screw in osteoporotic vertebral body;the group by pedicle screw augmented using calcium sulfate cement in osteoporotic vertebral body. Pedicle screw of equal specification was twisted into the tested pedicle of vertebral arch. The maximum axial screw pul-out strength and the maximum energy required to failure were recorded so as to assess the transient stability of pedicle screw augmented using calcium sulfate cement. RESULTS AND CONCLUSION:The maximum screw pul-out strength and the maximum energy required to failure were significantly less in osteoporotic vertebral body compared with normal vertebral body (P<0.05). The maximum screw pul-out strength and the maximum energy required to failure after augmentation using calcium sulfate cement were significantly increased (P<0.05). The maximum screw pul-out strength and the maximum energy required to failure after augmentation using calcium sulfate cement were identical between normal group and osteoporosis group. These results suggested that calcium sulfate cement could effectively increase the transient stability of pedicle screw. Calcium sulfate cement is effective in augmenting fixation in osteoporotic bone, and has potential in clinical application.

There are many different opinions about the taxonomy of plant pathogenic coryneform bacteria since they were departed from genus of Corynebacteria. In recent years, they were classified into 5 genus, including Clav-ibacter, Curtobacterium, Arthrobacter, Rhodococcus and Rathayibacter. Some new points of view about their taxonomy have been published thereafter. The changing of taxonomy is maily because of the methods'altering from old to new molecular and polyphasic taxonomy, and the latter is in continuously development. Taxonomy of plant pathogenic coryneform bacteria somehow depends on the cooperation of phytopathologists, microbilogists and other scientists.

Objective To investigate clinical characteristics and emergency managements of postrenal acute renal failure(ARV)induced by melamine in infant.Method Fluid therapy for urine alkalization and hydration,cistoscope drainage and peritoneal dialysis step by step were exerted in those who had both a history of certain milk intake and ARF according to the definition of pediatric ARF which developed by Pediatric Nephrology Assembly of Chinese Pediatric Association in 1994.Results Thirty-four postrenal ARF cases with anuria due to melamine in Nanjing Children's Hospital of Nanjing Medical University were involved in the study.Seventy cases(50%)re-ceived fluid therapy only.Nine cases(26.5%)received fluid thempy and eistoscope drainagemand 4 cases (11.8%)received fluid therapy and cistoscope drainage and peritoneal dialysis.Four cases(11.8%)received ur-gent peritoneal dialysis due to severe hyperkalemia.All cases(100%)survived.The urine pH at the first day.the second day,and after the second day in those who just pass away urine were 6.1±1.0、6.5±0.7.5.3±0.4,respectively(F=4.563,P=0.026).Conclusions Fluid therapy for urine alkalization and hydration and stop sequential thempy are effective in infant with postrenal ARF induced by melamine.

AIM:To explore the inhibitory effects of tumor associated mitochondrial protein 12 (TAMP12) on tumor cell apoptosis. METHODS: (1) A retrovirus expression vector was recombinated and transfected into the packaging cell line PA317. The virus particles were obtained to infect the target cell line HepG2 low expressing of TAMP12. The expression of TAMP12 mRNA was detected by RT-PCR. The subcellular localization and quantification of TAMP12 protein labeled with double fluorescein were observed under confocal laser scanning microscope (CLSM). (2) Hoechst33258 staining and flow cytometry (FACS) were used to analysis the apoptosis of HepG2 cells treated with 5-fluorouracil (5-FU). RESULTS: (1) The CLSM observation showed that TAMP12 protein was mainly expressed in mitochondria of HepG2 cells. The expressions of TAMP12 gene and protein were stable and high in transfected HepG2 cells. (2) Upon treatment with 5-FU, the transfected HepG2 cells showed a fairly integrated nucelus while the control HepG2 cells exhibited chromatin condensation, marginalization and karyorhexix. Moreover, the apoptosis rate of transfeced HepG2 cells was significantly lower than that in control HepG2 cells (P

Objectives To establish the two-dimensional electrophoresis(2-DE)profile of cell. Secreted proteins.Difierential expression profiling of fibroblast cell secreted proteins between nasopharyngeal carcinoma and normal nasopharyngeal tissue was analyzed.Methods Five tissue specimens each from patients with nasopharyngeal carcinoma and nasal polyp were collected individually.Fibroblast eells from above-mentioned tissue were cultured in serum-free medium,and cell-secreted proteins from the cultured medium were harvested by uhrafihration concentration and desalination.Samples were analyzed by 2-DE,and the differentially expressed proteins were analyzed and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry.Galectin-1 wa8 analyzed by EUSA test.Results 2-DE diagram of fibroblast cell-secreted proteins Was constructed.1 8 protein spots displayed quantitative changes in expression,and 11 protein spots among them were identified by mass speetrometrv.3 proteins including cystatin C,complement subcomponent C1S precursor,heterogeneous nuclear ribonueleoprotein A1 were down-regulated in the cultured medium of nasopharyngeal carcinoma associated fibroblast cells(CAFs). Nevertheless,the rest cell-secreted proteins including galectin-1,14-3-3 protein sigma,eathepsin L and etc,were up-regulated.Meanwhile,the expression of galectin-1 in the cultured medium was also analyzed and Its results were compared between CAFs and the normal fibroblast cells by ELISA.There Was statistical significance difference between them,and galectin-1 was up-regulated in the cIlltured medium of CAFs.Conclusions The changes of fibroblast cell-secreted proteins during nasopharyngeal carcinogenesi8 are analyzed by 2-DE analysis.The variation of pattern of secreted proteins is involved in signal transduction,protein synthesis,degradation and other pathways.CAFs may regulate tumor microenvironment by the abeve-mentioned pathways,and influence nasopharyngeal carcinogenesis,progress,invasion and metastasis.This study provided experimental basis for the eell secreted proteomics studv in future.

Objective To investigate curative effects of hypothermia on the secondary axotomy of nondisruptive axonal injury (NDAI) after diffuse brain injury (DBI).Methods A total of 16 male Sprague-Dawley rats were randomly and equally divided into hypothermia group (at 32℃ for 6 hours) and control group (at 37.5℃ ).The axonal swelling and axonal balls were detected by means of NF68kD immunochemistry after DBI caused by fluid percussion.The changes of maximal density of axonal swelling and axonal balls in callosum,diencephalon-mesencephalon,pons-oblongata and cerebellum were compared 24 and 72 hours after injury between both groups.Results NF68kD immunochemistry well showed axonal swellings and axonal balls in whole brain.The axonal swelling and axonal balls were significantly decreased 24 hours after DBI in both groups (P<0.05),especially in diencephalon-mesencephalon ,pons-oblongata and cerebellum (P<0.01).While there showed significant decrease of axonal swellings and axonal balls in pons-oblongata and cerebellum in hypothermia group 72 hours after DBI (P<0.05,P<0.01) but insignificant changes in the callosum and the diencephalon-mesencephalon compared with control group (P>0.05 ).Conclusions Hypothermia can retard the progress of mild or severe NDAI at early stage,which would taper with the longer time after injury except for partial mild NDAI.Hypothermia may prevent mild NDAI from secondary axotomy.

Biopsy, Fine-Needle ; methods ; Breast ; pathology ; Breast Neoplasms ; pathology ; Carcinoma, Ductal, Breast ; pathology ; Female ; Humans ; Immunohistochemistry ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Paraffin Embedding ; methods ; Receptor, ErbB-2 ; metabolism

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.