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.

Objective: To study the feasibility of percutaneous absorption of Compound Patch of Hyperosteogeny(CPH). Methods: The content of ferulic acid,an index composition in percutaneous receptor fluid and release receptor fluid were determined by HPLC.Results: The results of in vitro transdermal delivery experiment and in vitro release experiment showed that ferulic acid permeated at the constant speed of 0.2142?g?cm -2 ?h -1 in 24 hours and its release coincided with Higuchi Equation.Futhermore,the release rate was 14.53?g?cm -2 ?h -1/2 . Conclusion: CPH is a skeleton controlledtransdermal delivery system whose permeation speed is limited by skin.

Purpose:To study the effect of FasL gene on human lung adenocarcinoma cell lines and possibility of ex- ogenous FasL gene for gene therapy of lung cancer.Methods:An adenoviral expression vector with full length cDNA of FasL gene insert was constructed(Ad-FasL) and transfected into A549 cells.The effect of exogenous FasL gene on the growth of A549 cells was examined in vitro and in vivo.Results:Expression of FasL gene in A549 cells was confirmed by FCM and RT-PCR.The in vitro growth of the Ad-FasL transfected A549 cells was significantly inhibited (inhibition rate: 84%) as compared to mock (Ad-LacZ) transfected A549 cells.Colony-forming activity in vitro of the Ad-FasL transfected A549 cells was completely inhibited.The Ad-FasL transfected cells became apoptotic which was confirmed by the appear- ance of pre-G_1 on flow cytometry (FCM).The growth of A549 xenografts in nude mice was retarded by intra-tumol injection of Ad-FasL.Conclusions:FasL gene participates in the induction of cell apoptosis.Its use in gene therapy of cancer is promising.

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

Background and purpose:Promoter methylation ofPCDH10, a gene encoding protocadherin 10, has been found to be correlated to poor prognosis in gastric cancer (GC) patients. However, the relationship between the expression of PCDH10 and prognosis in GC remained unknown. This study aimed to explore the relationship be-tween the expression of PCDH10 and clinicopathological features and prognosis of GC, and to identify biomarker for predictions of recurrence and survival of GC.Methods:mRNA expressions of PCDH10 in 115 pairs of GC tissues and adjacent normal tissues were detected by real-time lfuorescence quantitative polymerase chain reaction (RTFQ-PCR). The correlation between PCDH10 expression level and clinicopathological features and prognosis of GC was analyzed. Prediction models for 5-year recurrence and 5-year survival were established using logistic regression method.Results:Progression-free survival (PFS) and overall survival (OS) were signiifcantly prolonged in patients with PCDH10 low expression compared to patients without PCDH10 low expression (P=0.046 andP=0.033 respectively). PCDH10 low expression signiifcantly correlated with less lymph node metastasis (P=0.001) and earlier TNM staging (P=0.001), and was more common in female than in male (P=0.040). The mRNA expression of PCDH10 did not correlate with age, Lauren classiifcation, T stage, neural invasion or vascular invasion. Univariate Cox analysis showed Lauren classiifca-tion, T stage, N stage, M stage and PCDH10 expression signiifcantly correlated with PFS and OS. Logistic regression models for the prediction of 5-year recurrence or 5-year survival based on clinicopathological features included Lauren classiifcation, T stage, N stage and M stage as variables. Logistic regression models for the prediction of 5-year recur-rence or 5-year survival based on PCDH10 expression included Lauren classiifcation, T stage, M stage and PCDH10 expression level but not N stage as variables. The models based on PCDH10 expression had the same effciencies as models based on clinical parameters in predicting 5-year recurrence or 5-year survival for GC patients.Conclusion:PCDH10 low expression correlated with better prognosis, less lymph node metastasis and earlier TNM stage in GC patients. Low expression of PCDH10 may be a biomarker of better survival for GC patients. Logistic regression model based on PCDH10 mRNA expression may serve as a prediction model when patients have unknown lymph node metas-tasis status.

OBJECTIVETo investigate the influence of Kudou Shencha decotion on INF-y, ICAM-1, MCP-1 levels of prostate tissue homogenate in immunity prostatitis model rats.

METHODForty Wistar male rats were divided into 5 groups randomly: Kudou Shencha decotion group with high dosage and low dosage, Qianleitai group, the model control group and normal group. The rat model of chronic nonbacterial prostatitis was established by multiple hypodermical injection of the suspension of prostatic protein purification with Freund's completed adjuvant. The level of intercellular adhesion molecule (ICAM-1), interferon gamma (INF-gamma) and monocyte chemotactic protein-1 (MCP-1) were measured by enzyme linked immunosorbent assay (ELISA).

RESULTThe content of ICAM-1 and MCP-1 in the model group was higher than that of the normal group (P < 0.05), the content of ICAM-1 was obviously decreased in Kudou Shencha decotion group with high dosage (P <0.05), the contents of MCP-1 were all obviously decreased in Kudou Shencha decotion groups and Qianlietai group. Compared with the model group, the contents of INF-gamma in all treatment groups were decreased insignificantly.

CONCLUSIONKudou Shencha decotion has the action of lowering the level of ICAM-1 and MCP-1, which may be one of the mechanisms of Kudou Shencha decotion in the therapy of chronic prostatitis.

Animals ; Chemokine CCL2 ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interferon-gamma ; metabolism ; Male ; Prostate ; drug effects ; metabolism ; Prostatitis ; drug therapy ; metabolism ; Rats ; Rats, Wistar

OBJECTIVE: To research the value of polymorphism of salivary esterase(Set) in paternity and personal identification. METHODS: Phenotype and genotype of human salivary esterase were detected in 114 liquid saliva samples from the Chinese population by disc electrophoresis and fast blue RR staining assay. RESULTS: The frequency of Set type was F 22.81%, FS 50.88%, S2 6.31%. The estimated gene frequency of SetF was 0.4825 and SetS was 0.5175. The PE was 0.1875 and the DP was 0.6199. CONCLUSION Polymorphism of salivary esterase (Set) was practical in paternity and personal identification.
Esterases/genetics* ; Forensic Anthropology/methods* ; Gene Frequency ; Humans ; Paternity ; Polymorphism, Genetic ; Saliva/enzymology*

Objective To improve the biological fidelity of the thorax flexible body in the original MADYMO child human model,so as to further study pediatric thorax injuries of child occupant.Methods The finite element model of six-year-old pediatric thorax was built by the method of reverse modeling based on CT images.By replacing the thorax model with flexible body in MADYMO six-year-old human model,an improved human model containing biomechanical thorax model was developed.The model was verified by joint validation of two tests,including Irwin and Mertz's method of scaling channel reported in Kroell's adult chest impact experiment and Ouyang's thoracic impact test on pediatric cadavers.Results The response of this established thorax model was in good agreement with scaling channel method and cadaver test data,and the thorax model was much more accurate than the original flexible body model.The resilience of simulation model was consistent with cadaver test.Conclusions The validity of the model is verified,and the results can be further used for occupant injury analysis in vehicle frontal crash.

BACKGROUND: Particulated juvenile cartilage allograft is simple and easy to obtain, and relevant clinical studies are underway in the USA. However, how the transplanted juvenile cartilage fragments exert biological effects through biochemical mechanisms and genetic signal transduction is still unclear. There is as yet no report on this technology in China. OBJECTIVE: To explore the feasibility of articular cartilage defects repaired with particulated juvenile cartilage allograft. METHODS: The cartilage fragments were obtained from juvenile Pitman-Moore strains (provided by the Laboratory Animal Center of the Army Medical University in China) and cultured in vitro. Brdu immunofluorescence assay was performed at 1, 3, and 7 days of culture. The particulated juvenile cartilage allograft/fibrin gel composites were subcutaneously transplanted into the SCID rats (provided by the Laboratory Animal Center of the Army Medical University). The specimens were taken for hematoxylin-eosin staining, safranin O staining and immunohistochemistry after 1 month. Cartilage defects of 8 mm in diameter were made in the knee joint of 10 adult Pitman-Moore strains (Laboratory Animal Center of the Army Medical University), and were randomized into two groups, which were then transplanted with the particulated juvenile cartilage allograft/fibrin gel composites (experimental group) or nothing (control group). The specimens were taken for hematoxylin-eosin staining, safranin O fast green staining, toluidine blue and immunohistochemistry at 3 months after transplantation. RESULTS AND CONCLUSION: Little Brdu incorporation was detected in juvenile cartilage fragments at 1 day of culture, some Brdu incorporation was defected at 3 days of culture. At 7 days of culture, a progressive increase in the Brdu signal was detected in chondrocytes within the cultured cartilage fragments, which seemed to localize along the tissue edge. At 1 month after subcutaneous transplantation, the particulated juvenile cartilage allograft still survived and were surrounded by few proliferative chondrocytes. There was no obvious tissue repair in the control group at 3 months after transplantation. In the experimental group, there was obvious tissue repair, the color of the newly formed tissues was similar to the normal cartilage tissue, which integrated well with the surrounding normal cartilage tissue, and the cells distributed evenly. These results imply that particulated juvenile cartilage allograft can achieve good results in repairing articular cartilage defects.

Aim To observe the effects of realgar nano-particles on B cell non-Hodgkin's lymphoma Raji cells in vitro. Methods Realgar nanoparticles and crude realgar particles were characterized with a laser particle size analyzer, a transmission electron microscopy (TEM)and an atomic force microscopy(AFM). The morphological changes of proliferation of Raji cells brought about by the use of realgar naoparticles and crude realgar particles were observed with a light mi-croscope. The membrane changes of Raji cells treated with realgar naoparticles and crude realgar particles were observed with AFM. The ultrastructures of Raji cells were observed with TEM. The inhibitory effects of Raji cells treated with realgar naoparticles and crude realgar particles were measured with MTT. The nuclear apoptosis morphologies of Raji cells were observed with fluorescence microscopy. The apoptosis rates and the cell cycle distributions of Raji cells treated with real-gars were measured with flow cytometry. Results The size of realgar nanoparticles and crude realgar particles was (79 ± 8)nm and (1. 89 ± 0. 2)μm,respectively. Light microscopy showed that realgar nanoparticles could inhibit the aggregation growth of Raji cells. AFM showed that Raji cells treated with realgar nanoparticle became shrank, had smaller volume and lost the growth state of stretching out. Raji cells treated with crude realgars did not change significantly. TEM showed Raji cells treated with realgar nanoparticle had damaged subcellular organelles and mitochondria with increased vacuoles. The Raji cells treated with crude realgar did not change significantly. MTT assay showed that when treated with the final concentration of 50 mg ·L - 1 of realgar nanoparticle for 24 h,the cell survival rate of Raji cells was (40 ± 2)% . When treated with the same concentration of crude realgar,its survival rate was (65 ± 3)% . When treated with 50 mg·L - 1 of realgar nanoparticle for 48 h,its survival rate was only 10 % ,and when treated with crude realgar ,its survival rate was (42 ± 2 )% . Fluorescence micro-scope indicated that the Raji cells treated with realgar nanoparticle had obvious nuclear apoptosis,which was not obvious in crude realgar group. Flow cytometry showed that the total apoptosis rate of Raji cells in-duced by realgar nanoparticles and by crude realgar was 11. 14%,15. 9%,respectively. Compared with those treated with crude realgar,the Raji cells treated with realgar nanoparticles presented a significantly higher ratio cell distribution in G1 phase and an obvious decreased ratio in S phase. Conclusion Compared with crude realgar particles,the same dose of realgar nanoparticles can significantly inhibit the proliferation of Raji cells,destroy their sub-cellular structure,and induce the cell apoptosis of Raji cells.