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.

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.

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.

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.

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*

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

OBJECTIVETo study the effect of Hewei Xiaopi Capsule (HXC) in treating patients with dyskinesis functional dyspepsia (FD).

METHODSSixty-three patients with FD were randomly assigned to two groups, 33 in the treated group and 30 in the control group, who were treated respectively with HXC and domperidone for 4 weeks to observe the clinical effect and changes in electrogastrogram (EGG) before and after treatment.

RESULTSSymptoms were alleviated in both groups after treatment, 3 patients in the treated group were cured, the efficacy was judged as markedly effective in 11, effective in 14 and ineffective in 5, while in the control group, 1 cured, 8 markedly effective, 14 effective and 7 ineffective, difference between the two groups showed no statistical significance. EGG showed that in the treated group, the normal slow-wave percentage was 41.93 +/- 18.22 before treatment and 50.86 +/- 16.03 after treatment, showing significant difference (P < 0.05).

CONCLUSIONHXC is markedly effective in treating patients with FD.

Drugs, Chinese Herbal ; therapeutic use ; Dyskinesias ; drug therapy ; Dyspepsia ; drug therapy ; Gastrointestinal Agents ; therapeutic use ; Humans ; Tablets

OBJECTIVETo compare the relation between the preoperative functional magnetic resonance imaging (fMRI) with blood oxygen level dependent (BOLD) technique and intraoperative motor evoked potential (MEP) monitoring for cortical mapping of primary motor cortex in patients with tumors near the central area. And to determine whether non-invasive preoperative fMRI can provide results equivalent to those achieved with the invasive neurosurgical "gold standard".

METHODSA prospective study of 16 patients with various pathological tumors of the central area was conducted. Preoperative fMRI scans using the BOLD contrast technique in each patient were performed. An activation scan was achieved by using a motor task paradigm, which consisted of simple flexion-extension finger movements and finger-to-thumb touching in a repeating pattern. The anatomical structure was delineated by the T(1)-weighted three-dimensional fast spoiled gradient recalled sequence (3D/FSPGR) immediately afterward. The BOLD images were overlaid on the T(1)-weighted 3D/FSPGR images, and then co-registered to the neuronavigation system. The fMRI activations were documented by using a neuronavigation system in sequence, and compared to standardized intraoperative MEP monitoring, which included direct cortical electrical stimulation (DCES) or transcranial cortical electrical stimulation (TCES) or their combination. The compound muscle action potentials of forearm flexor and hand muscle responses were recorded during either TCES or DCES. Two techniques were compared to determine the accuracy for cortical mapping of primary motor areas with fMRI.

RESULTSOverall, the intraoperative MEP monitoring showed good correlation with fMRI activation in 92.3% of cases. The coincidence rate, however, was 100.0% between TCES and fMRI, and 66.7% between DCES and fMRI respectively. There was no statistically difference between two cortical mapping techniques, chi-square test of paired comparison of enumeration data, P < 0.01.

CONCLUSIONBOLD fMRI was a high sensitive and reliable technique to locate the position of the primary motor areas and their spatial relation with adjacent tumor, especially for the presurgical planning in patients with central area brain tumor.

Adolescent ; Adult ; Brain Neoplasms ; pathology ; physiopathology ; surgery ; Child ; Evoked Potentials, Motor ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Monitoring, Intraoperative ; Motor Cortex ; pathology ; physiology ; Neuronavigation ; Oximetry ; Prospective Studies ; Transcranial Magnetic Stimulation ; methods