Association of Haplotypes in Exon 4 of KLK2 Gene with Raised Serum Prostate-Specific Antigen

The standard diagnostic modalities for Prostate Cancer (PC) include serum Prostate-Specific Antigen (PSA) assay, Digital Rectal Examination (DRE), and histological examination of prostate biopsy. They are limited by low predictive potential and inability to predict which patients are at risk of developing metastatic disease. The aim of this study is to investigate the exon 4 of the KLK2 gene of subjects for changes in its nucleotide sequences (SNPs) and determine the correlation of these changes with serum PSA in an Igbo population of Nigeria. One hundred male subjects aged 40 years and above, who gave their consent, were used for the study. Their PSA determinations were done using ELISA technique while genetic studies were carried out using real-time PCR. tPSA, fPSA, and % fPSA of the subjects ranged between 0.8% - 18.30%, 0.10% - 1.60% and 0.0% - 0.7% respectively. Of the 100 subjects, 28 subjects had tPSA levels above 4.0 ng/ml with a mean of 7.10 (±3.30) ng/ml. Those with tPSA less than 4 ng/ml had a mean of 1.87 (±0.85) ng/m. 15 subjects showed SNPs with a mean tPSA of 6.87 (±4.82) ng/ml while the remaining 85 subjects without SNPs had a mean of 1.86 (±0.80) ng/ml. Results from direct DNA sequencing showed 11 SNPs. Ten subjects are curated in SNP database while one is uncurated. The Chi-square test showed significant association (p = 0.00) between tPSA levels and SNPs mutation (X2 = 17.35, p = 0.00). A Kruskal-Wallis test demonstrated that the positional arrangement of the SNP mutations had no effect on PSA-total or free-values (H (10) = 10.92, p = 0.28;H (10) = 10.07, p = 0.38 respectively). Two SNPs: rs6072 and rs74478031 were associated with elevated PSA levels (p < 0.05). Their presence, therefore, has the potential to serve, in conjunction with raised PSA, as biomarkers of prostate cancer in the study population.

Prostate-specific antigen half-life： a new predictor of progression- free survival and overall survival in Chinese prostate cancer patients

We investigated the potential value of prostate-specific antigen half-life （PSAHL） and decreasing velocity （PSAVd） to predict progression-free survival （PFS） and overall survival （OS） in Chinese patients with prostate cancer. A total of 153 patients treated with hormonal therapy were included in the study. Of these, 78 patients progressed to hormone- refractory prostate cancer （HRPC） and 24 patients died by the end of follow-up. PSAHL was defined as the time during which prostate-specific antigen （PSA） concentration became half of the initial value during the first hormonal therapy. PSAVd reflected the decreasing velocity of PSA during the first hormonal therapy. PFS was defined as the interval from the beginning of hormonal therapy to HRPC. Cox proportional hazards regression analysis was used to evaluate whether PSAHL and PSAVd were significantly associated with PFS and OS. The median PSAHL and PSAVd were 0.50 months and 33.8 ng mL^-1 per month. The median PFS and OS were 22.7 months （95% confidence interval [CI], 22.0-29.6 months） and 43.5 months （95% CI, 37.9-48.4 months）, respectively. On univariate and multivariate analysis, long PSAHL （〉 0.5 months）, metastatic disease, high biopsy Gleason scores （〉 8） and high nadir PSA （〉 0.4 ng mL^-1） were all found to be significantly associated with short PFS. Long PSAHL, high nadir PSA and short PSA doubling time （PSADT 〈 2.0 months） were significantly associated with short OS. There were no significant relationships between PSAVd and either PFS or OS. Thus, PSAHL is a promising new independent predictor of survival. Patients with long PSAHL were identified as those at high risk for a relatively short PFS and OS.

Tumor-Specific Histo-Blood Group Antigens: Apropos of Two Cases

Cancer cells with immunogenic properties having altered protein glycosylation, modified blood group substances have been widely studied. Due to the genetic instability occurring during carcinogenesis the glycosyltransferases may suffer from posttranslation sequence modification. The author describes 2 autopsy cases, where in the background of the unusual metastatic tumor presentation, incompatible blood group antigenic determinants have been demonstrated using blood group specific lectins and monoclonal antibodies (mAb). In the first case, reported here, a 10-year-old girl developed an acute myeloid leukemia and died in a septic endotoxin shock after successful cytostatic treatment of a juvenile signet ring cell cancer of her colon. At autopsy there were no signs of tumor except bilateral apple-sized mucinous ovarian (Krukenberg) metastases. While she had erythrocyte phenotype of blood group A, the signet ring adenocarcinoma cells expressed blood group B incompatible antigenic determinants with lectin/mAb. In the second case, the autopsy of a 78-year-old female resulted in no macroscopic tumor sign except a moderately enlarged, ham hard spleen. Light microscopy revealed adenocarcinomatous infiltration in the splenic sinusoids. The patient had blood group O, while the metastatic cells in the spleen reacted with Breast Carcinoma Antigen (BioGenex) and incompatible anti-B Banderiaeasimplicifolia agglutinin I and anti-B mAb. It proved to be a case of an occult, completely regressed breast cancer. Based on these observations the expression of tumor specific incompatible blood group antigens might occur from time to time, mostly in adenocarcinomas. Accordingly, blood group-based specific immuno-oncotherapy could be considered in some cancer cases.

Decreasing trend in prostate cancer with high serum prostate-specific antigen levels detected at first prostate-specific antigen-based population screening in Japan

To clarify the recent trends in prostate-specific antigen （PSA） distribution in men in Japan, we analyzed the PSA distributions of men undergoing PSA-based population screening. We summarized the annual individual data of PSA-based population screening in Kanazawa, Japan, from 2000 to 2011, and analyzed baseline serum PSA values of the participants at the first population screening. Serum PSA distributions were estimated in all participants and those excluding prostate cancer patients according to age. From 2000 to 2011, 19 620 men participated aged 54-69 years old in this screening program. Mean baseline serum PSA level of all participants at the first screening was 2.64 ng m1-1 in 2000, and gradually decreased to approximately 1.30 ng ml-I in 2006. That of participants excluding prostate cancer patients was 1.46 ng m1-1 in 2000, and there was no remarkable change during the study period. The 95t＂ percentiles in the participants excluding prostate cancer patients detected at the first population screening of men aged 54-59, 60-64, and 65-69 years old were 2.90, 3.60, and 4.50 ng m1-1, respectively. After the commencement of population screening, the proportion of prostate cancer patients with high serum PSA levels decreased. However, there were no changes in serum PSA levels in men without prostate cancer. Age-specific PSA reference level of men without prostate cancer in Japan was similar to that in China and Korea.

Body mass index and serum lipid profile influence serum prostate-specific antigen in Chinese men younger than 50 years of age

This study is to assess the potential factors that could affect the serum prostate-specific antigen （PSA） level in healthy younger men. We evaluated the associations of age, body mass index （BMI） and serum lipid profile with serum PSA level in 6774 Chinese men （aged 20-49 years） who received a routine health examination. Eligible men were classified into 10-year age groups, BMI was categorized as underweight （〈18.5）, normal （18.5-22.9）, overweight （23.0-24.9）, obese （25.0-29,9） and very obese （〉30） according to the redefined World Health Organization （WHO） criteria for the Asia-Pacific region. PSA levels were compared among groups as well, In multiple linear regression analysis, PSA was positively correlated with age （P〈0.0001）. Negative correlations existed between PSA and BMI （P〈0.0001） and triglyceride level （P=0.01）. No relationship could be found between PSA and serum cholesterol （P=0.711） or high-density lipoprotein （HDL; P =0.665）. In addition, we found that serum PSA levels increased with age and decreased with BMI. Our study demonstrates that age, BMI and triglyceride levels influence the PSA level in men 〈50 years of age.

Radioiodine therapy for castration-resistant prostate cancer following prostate-specific membrane antigen promoter-mediated transfer of the human sodium iodide symporter

Radioiodine therapy, the most effective form of systemic radiotherapy available, is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter （hNIS）. Here, we explore the efficacy of a novel form of gene therapy using prostate-specific membrane antigen （PSMA） promoter-mediated hNIS gene transfer followed by radioiodine administration for the treatment of castration-resistant prostate cancer （CRPC）. The androgen-dependent C33 LNCaP cell line and the androgen-independent C81 LNCaP cell line were transfected by adenovirus. PSMA promoter-hNIS （Ad.PSMApro-hNIS） or adenovirus.cytomegalovirus-hNIS containing the cytomegalovirus promoter （Ad.CMM-hNIS） or a control virus. The iodide uptake was measured in vitro. The in vivo iodide uptake by C81 cell xenografts in nude mice injected with an adenovirus carrying the hNIS gene linked to PSMA and the corresponding tumor volume fluctuation were assessed. Iodide accumulation was shown in different LNCaP cell lines after Ad.PSMApro-hNIS and Ad.CMV-hNIS infection, but not in different LNCaP cell lines after adenovirus.cytomegalovirus （Ad.CMV） infection. At each time point, higher iodide uptake was shown in the C81 cells infected with Ad.PSMApro-hNIS than in the C33 cells （P 〈 0.05）. An in vivo animal model showed a significant difference in 1311 radioiodine uptake in the tumors infected with Ad.PSMApro-hNIS, Ad.CMV-hNIS and control virus （P 〈 0.05） and a maximum reduction of tumor volume in mice infected with Ad.PSMApro-hNIS. These results show prostate-specific expression of the hNIS gene delivered by the PSMA promoter and effective radioiodine therapy of CRPC by the PSMA promoter-driven hNIS transfection.

A nomogram based on age, prostate-specific antigen level, prostate volume and digital rectal examination for predicting risk of prostate cancer

Nomograms for predicting the risk of prostate cancer developed using other populations may introduce sizable bias when applied to a Chinese cohort. In the present study, we sought to develop a nomogram for predicting the probability of a positive initial prostate biopsy in a Chinese population. A total of 535 Chinese men who underwent a prostatic biopsy for the detection of prostate cancer in the past decade with complete biopsy data were included. Stepwise logistic regression was used to determine the independent predictors of a positive initial biopsy. Age, prostate-specific antigen （PSA）, prostate volume （PV）, digital rectal examination （DRE） status, % free PSA and transrectal ultrasound （TRUS） findings were included in the analysis. A nomogram model was developed that was based on these independent predictors to calculate the probability of a positive initial prostate biopsy. A receiver-operating characteristic curve was used to assess the accuracy of using the nomogram and PSA levels alone for predicting positive prostate biopsy. The rate for positive initial prostate biopsy was 41.7% （223/535）. The independent variables used to predict a positive initial prostate biopsy were age, PSA, PV and DRE status. The areas under the receiver-operating characteristic curve for a positive initial prostate biopsy for PSA alone and the nomogram were 79.7% and 84.8%, respectively. Our results indicate that the risk of a positive initial prostate biopsy can be predicted to a satisfactory level in a Chinese population using our nomogram. The nomogram can be used to identify and counsel patients who should consider a prostate biopsy, ultimately enhancing accuracy in diagnosing prostate cancer.

Prostate-specific antigen kallikrein and the heart

Currently,there is growing interest regarding prostatespecific antigen(PSA) and the cardiovascular system.Increased PSA serum levels have been reported after prolonged cardiopulmonary resuscitation,cardiac surgery,extracorporeal cardiopulmonary bypass,acute myocardial infarction(AMI) and coronary artery stenting.The possible role of PSA in cardiac events has been questioned due to the finding of PSA decrease during AMI and by the correlation of variation in PSA levels with coronary lesions and occurrence of major adverse cardiac events.Complexed PSA forms and uncomplexed PSA forms are observed in the bloodstream but the increasing formation of irreversible bound PSA seems to be a crucial finding during AMI.Large studies need to be carried out to confirm these preliminary results and to elucidate unclear aspects.These findings present many potential directions for future research including the role of uncomplexed forms of PSA,the possible distribution of PSA in the heart,the relative expression levels in heart disease states,the mode of expression regulation and other potential specific substrates.The journey of PSA investigation could be longer than initially expected.

Serum prostate-specific antigen as a predictor of prostate volume and lower urinary tract symptoms in acommunity-based cohort： a large-scale Korean screening study

The aim of this study is to assess the ability of serum prostate-specific antigen （PSA） to predict prostate volume （PV） and lower urinary tract symptoms （LUTS） represented by the international prostate symptom score （IPSS）. From January 2001 to December 2011, data were collected from men who first enrolled in the Korean Prostate Health Council Screening Program. Patients with a serum PSA level of 10 ng ml^-1 or age 〈40 years were excluded. Accordingly, a total of 34 857 men were included in our study, and serum PSA, PV and the IPSS were estimated in all patients. Linear and age-adjusted multivariate logistic analyses were used to assess the potential association between PSA and PV or IPSS. The predictive value of PSA for estimating PV and IPSS was assessed based on the receiver operating characteristics-derived area under the curve （AUC）. The mean PV was 29.9 ml, mean PSA level was 1.49 ng ml^-1 and mean IPSS was 15.4. A significant relationship was shown between PSA and PV, and the IPSS and PSA were also significantly correlated after adjusting by age. The AUCs of PSA for predicting PV ~20 ml, 〉25 ml and 〉35 ml were 0.722, 0.728 and 0.779, respectively. The AUCs of PSA for predicting IPSS 〉 7, 〉 13 and 〉 19 were 0. 548, 0.536 and 0. 537, respectively. Serum PSA was a strong predictor of PV in a community-based cohort in a large-scale screening study. Although PSA was also significantly correlated with IPSS, predictive values of PSA for IPSS above the cutoff levels were not excellent. Further investigations are required to elucidate the exact interactions between PSA and LUTS and between PSA and PV in prospective controlled studies. Such studies may suggest how PSA can be used to clinically predict PV and the IPSS.

Relationship between insulin resistance, obesity and serum prostate-specific antigen levels in healthy men

The purpose of this study was to determine the relationship between insulin resistance, obesity and serum prostate-specific antigen （PSA） levels in healthy men with serum PSA level below 4 ng mL-1. The men included in the study cohort were 11 827 healthy male employees of the Korea Hydro and Nuclear Power Co., LTD who had undergone medical checkups including fasting glucose, fasting insulin and serum PSA between January 2003 and December 2008. Insulin resistance was calculated by homeostasis model assessment （HOMA [fasting glucose × fasting insulin]/22.5） and quantitative insulin sensitivity check index （QUICK/; 1/[log （fasting insulin） ＋ log （fasting glucose）]）. Age-adjusted body mass index （BMI） was significantly increased according to increasing quartile of insulin resistance as determined by HOMA and QUICKI, respectively, in analysis of variance （ANOVA） test and Duncan＇s multiple comparison test （P 〈 0.001）, but age-adjusted serum PSA concentration was significantly decreased according to increasing quartile of insulin resistance as determined by HOMA and QUICK/（P 〈 0.001）. Age, BMI, insulin resistance by HOMA or QUICK/were significantly independent variables to serum PSA level in a multivariate linear regression analysis （P 〈 0.001）. Insulin resistance was a significant independent variable to serum PSA level along with BMI. Insulin resistance and BMI were negatively correlated with serum PSA level in healthy men. Insulin resistance was positively correlated with BMI.

A population study of fasting time and serum prostate-specific antigen （PSA） level

Prostate cancer is one of the most common cancers in men. Traditional screening and diagnostic methods include digital rectal examinations （DREs）, biopsies and serum prostate-specific antigen （PSA） tests, with the latter being the more popular. PSA is a biomarker for prostate cancer; however, it is highly sensitive to external factors as well as other prostate diseases. As such, the reliability of of the serum PSA level as a sole screening and diagnostic tool for prostate cancer is controversial. Recently, it has been shown that fasting extremes can affect concentrations of serum chemistry analytes, thus raising the question of whether or not fasting has an effect on the highly sensitive PSA biomarker. Patients testing for serum PSA levels are often concomitantly submitting to other tests that require fasting, subjecting certain patients to a fasting PSA level while others not. The objective of this study was to investigate whether this discrepancy in fasting state translates into an effect on serum PSA levels. Serum PSA levels and fasting time records for 157 276 men who underwent testing at Calgary Laboratory Services （CLS; Calgary, Alberta, Canada） between 01 January 2010 and 31 March 2013 were accessed. Linear regression models of mean PSA levels and fasting times revealed a statistically important relationship at certain fasting times. Applying a dynamic mathematical model to explore the clinical effect of fasting suggests minimal impact on serum PSA result interpretation. Thus, patients can be tested for serum PSA levels regardless of their fasting state.

Salvage treatments for prostate-specific antigen relapse of cT3N0M0 prostatic adenocarcinoma after radical prostatectomy combined with neoadjuvant androgen deprivation

Objective The aim of the study was to evaluate the efficiency of salvage treatments for prostate specific antigen(PSA)relapse of cT3N0M0 prostatic adenocarcinoma(PCa)after radical prostatectomy(RP)combined with neoadjuvant androgen deprivation(ADT).Methods A total of 332 patients with cT3N0M0 PCa were enrolled in the prospective study and received RP and pelvic lymph node dissection with neoadjuvant ADT for 3 months.All patients with PSA relapse were treated with salvage external beam radiation therapy(RT)and ADT for 6 months.Results The 5-year postoperative PSA relapse rate was 40.96%(136/332).The patients have been divided into the PSA relapse and PSA relapse-free groups in order to compare patient characteristics.The ratio of patients with Gleason score≥8 and positive surgical margin in the PSA relapse group were significantly higher than those of in the PSA relapse-free group(P=0.01).The mean duration between the start of operative treatment and PSA relapse was 31 months.Salvage treatment to all 136 PSA relapse patients led to favorable outcomes.PSA relapse was not observed after salvage treatment by the end of follow-up.The 5-year overall survival rates of the PSA relapse and PSA relapse-free groups were 94.9%and 93.9%,respectively.Conclusion In pursuit of curative treatment,our study showed that RP combined with neoadjuvant ADT is an aggressive multimodality strategy associated with lower PSA relapse and better survival outcomes for stage cT3N0M0 PCa patients.Patients with PSA relapse after RP may benefit from early aggressive salvage RT combined with short-term ADT.

Prostate-specific antigen doubling time and response to cabazitaxel in a hormone-resistant metastatic prostate cancer patient

We report a case of metastatic castration-resistant prostate cancer, who received prior treatment with docetaxel and was then given cabazitaxel as salvage therapy. The patient was monitored by prostate-specific antigen doubling time and prostate-specific antigen absolute value. The prostate-specific antigen doubling time was found to be a good response predictor in the patient.

Alternative mechanisms for prostate-specific antigen elevation:A prospective analysis of 222 transurethral resections of prostate patients

AIM: To investigate the relationship between prostatespecific antigen(PSA) levels and(1) bladder outlet obstruction(BOO) and(2) the severity of prostate inflammation.METHODS: Two hundred and twenty-two consecutive patients undergoing transurethral resection of the prostate(TURP) were prospectively included. Patients with proven urinary tract infection and/or known prostate cancer were excluded. PSA levels, International Prostate Symptoms Score(IPSS), prostate weight, post residual volume and pressure flow parameters were determined. A histopathological assessment of the presence and severity of inflammation was also performed.RESULTS: Patients had a mean age of 69.1 ± 8.6 years(45-90 years), with mean preoperative PSA levels of 4.7 ± 5.4 ng/m L(0.2-32.5 ng/m L) and IPSS of 15.7 ± 6.9(0-32). Mean Pdet Q max was 96.3 ± 34.4 cm H2O(10-220 cm H2O). The mean resected prostate weight was 39.4 ± 27.3 g(3-189 g). Correlations were observed between PSA(logarithmic) and resected prostate weight(r = 0.54; P < 0.001), PSA(logarithmic) and Pdet Q max(r = 0.17; P = 0.032), and resected prostate weight and Pdet Q max(r = 0.39; P < 0.001). Furthermore, low correlations were observed between PSA(logarithmic) and active(r = 0.21; P < 0.0001) and chronic(r = 0.19; P = 0.005) inflammation. CONCLUSION: In this study we showed a correlation between BOO(Pdet Q max) and PSA(logarithmic). Furthermore, we demonstrated a weak correlation between PSA(logarithmic) and active as well as chronic prostatic inflammation.

Total and free prostate-specific antigen indexes in prostate cancer screening:value and limitation for Japanese populations

Aim:To assess the efficacy and limitation of free/total prostate-specific antigen ratio(f/tPSA)at a single institution in Japan,focusing on the avoidance of pointless prostate biopsies.Methods:In total,631 men between 44 and 93 years old(mean 69.8 years)with elevated PSA underwent power-Doppler ultrasoundgraphy-guided transrectal 10-core prostate biopsies at Niigata Cancer Center Hospital,and their histological features were investigated with total PSA (tPSA)and f/tPSA.Results:PCa was detected in 126 of 134 patients(94.3%)with tPSA of 26 ng/mL or higher.The detection rate was 59.4% for tPSA of 21-25 ng/mL,followed by 39.2% for 16-20 ng/mL,30.0% for 11-15 ng/mL, 20.0% for 4.1-10 ng/mL and 7.6% for≤4.0 ng/mL,f/tPSA of the PCa group was significantly lower than that of non-malignamt disorders in any tPSA ranges(mean 0.122 vs.0.160,P<0.001).Receiver-operating characteristics analyses showed that f/tPSA(AUC:0.664)performed more valuably than tPSA(AUC:0.559)in patients with tPSA between 3.0-10 ng/mL(P<0.01).Although f/tPSA of 0.250 for the cut-off value might miss 1.8% PCa patients,it potentially spares 9.2% of unnecessary biopsies.Conclusion:f/tPSA is more valuable compared with tPSA alone for the prediction of the occurrence of PCa.We recommend 0.250 as the cut-off value for f/tPSA in PCa screening for Asian men having so-called grey-zone tPSA.(Asian J Androl 2006 Jul;8:429-434)

Neuron specific enolase,p53蛋白和proliferating-cell nuclear antigen在肺癌组织中的表达及意义

目的：研究神经元特异性烯醇化酶（neuronspecificenolase，NSE）的表达，与p53蛋白和增殖细胞核抗原（prolif－erating－cellnuclearantigen，PCNA）表达的关系及意义．方法：用特异性鼠抗人单克隆抗体，按LSAB免疫组织化学方法检测石蜡包埋的肺癌标本中NSE，p53蛋白和PCNA的表达．结果：在小细胞性肺癌（SmallCellLungCancer，SCLC）中，NSE阳性表达者，PCNA标记指数（LabellingIndex，LI）高于NSE阴性者，而p53蛋白的表达与NSE的表达无关．在非小细胞性肺癌（non－smallcelllungcancr，non－SCLC）中，p53蛋白的表达和PCNALI均与NSE的表达无关．结论：在SCLC的发生及发展过程中，神经内分泌功能可能起了部分作用．而p53抑癌基因在SCLC的发生中并不起着重要的作用．

Mass screening of prostate cancer in a Chinese population:the relationship between pathological features of prostate cancer and serum prostate specific antigen

Aim:To investigate the pathological features of the prostate biopsy through mass screening for prostate cancer in a Chinese cohort and their association with serum prostate specific antigen (PSA).Methods:A total of 12 027 Chinese men in Changchun were screened for prostate cancer by means of the serum total prostate specific antigen (tPSA) test (by Elisa assay).Transrectal ultrasound-guided systematic six-sextant biopsies were performed on those whose serum tPSA value was >4.0 ng/mL and those who had obstructive symptoms (despite their tPSA value) and were subject to subse- quent pathological analysis with the aid of the statistic software SPSS 10.0 (SPSS.Inc.,Chicago.USA).Results:Of the 12 027 cases,158 (including 137 patients whose serum tPSA values were >4.0 ng/mL and 21 patients [serum tPSA <4. 0 ng/mL] who had obstructive symptoms) undertook prostate biopsy.Of the 158 biopsies,41 cases of prostatic carci- noma were found (25.9 %,41/158).The moderately differentiated carcinoma and poorly differentiated carcinoma ac- counted for 61% and 34 %,respectively.A significant linear positive correlation between the serum tPSA and the Gleason scores in the 41 cases of prostatic carcinoma (r=0.312,P<0.01) was established.A significant linear positive correlation between the serum tPSA value of the 41 prostatic carcinoma and the positive counts of carcinoma in sextant biopsies was established (r=0.406,P<0.01),indicating a significant linear relationship between serum tPSA and the size of tumor. Condusion:This study was the first to conduct mass screening for prostate cancer by testing for serum tPSA values and the first to investigate the pathological features of prostate cancer in a cohort of Chinese men.Our results reveal that the moderately differentiated carcinoma is the most common type of prostate cancer.This study also has shown that the serum tPSA value in prostate cancer is associated with the Gleason score and the size of tumor.

Analysis of Protoscoleces-specific Antigens from Echinococcus Granulosus with Proteomics Combined with Western Blot

Objective To establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research. Methods Brood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer＇s instructions. We employed two-dimensional electrophoresis （2-DE） combined with immunoblot assay （Western blot） to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software. Results About 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16 000 Da to 117 000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified. Conclusion 2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.

INTRATHYMIC INOCULATION OF LIVER SPECIFIC ANTIGEN ALLEVIATES LIVER TRANSPLANT REJECTION

Objective To study the effects of liver specific antigen(LSA)on liver allotransplantation rejection. Methods Orthotopic liver transplantation was performed in this study. GroupⅠ: syngeneic control(Wistar-to-Wistar); GroupⅡ: acute rejection(SD-to-Wistar). GroupIII: thymic inoculation of SD rat LSA day 7 before transplantation. The observation of general condition and survival time, rejection grades and the NF-кB activity of splenocytes were used to analyze severity of acute rejection and immune state of animals in different groups. Results The general condition of groupⅠwas fair post transplantation with no sign of rejection. All recipients of group Ⅱ died within days 9 to 13 post transplantation with median survival time of 10.7 ±1.37 days. As for group III, 5 out of 6 recipients survived for a long period with remarkably better ge-neral condition than that of group Ⅱ. Its rejection grades were significantly lower than groupⅡ(P < 0.05).NF-кB activity was only detected in groupⅠbetween days 5 and 7 after transplantation, whereas highactivity of NF-кB was detected at all points in group Ⅱ and low NF-кB activity was detected in group III which was significantly lower than that of group Ⅱ(P < 0.05). Conclusions LSA is an important transplantation antigen directly involved in the immunorejection of liver transplantation. Intrathymic inoculation of LSA can alleviate the rejection of liver allotransplantation,grafts survive for a period of time thereby, allowing a novel way to liver transplantation immunotolerance.

Detection of human liver-specific F antigen in serum and its preliminary application

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