Objective To evaluate the false-positive of Abbott chemiluminescence immunoassay (CLIA) in HIV screening test. Methods The total 39076 samples from the pre-operation patients (from August 2011to March 2012) in Beijing Tongren Hospital were tested for HIV screening through the ARCHITECT I2000. Results 31samples were screened as the anti-HIV positive.After the HIV confirmation test by Beijing Center for Disease Control, 21samples were diagnosed as the anti-HIV positive, 6 samples were indeterminate and 4 samples were anti-HIV negative. Conclusion The false-positive rate of ARCHITECT HIV Ag/Ab Combo is high(12.90%), the anti-HIV positive should be diagnosed based on clinical data and HIV confirmation test with regular follow-up.

为探讨缺血性脑卒中患者ET、CGRP及血清MDA、SOD测定的意义，用放射免疫分析法及TBA比色法测定了患者血浆ET、CGRP及血清MDA、SOD水平。 结果提示，小面积梗死组和大面积梗死组ET水平均显著高于正常对照组（P 均小于0.01)。两病理组之间比较均存在显著差异(P 均小于0.05)。大面积组ET与CGRP呈显著负相关(r =-0.668，P＜0.05)。MDA及SOD统计值的变化关系与ET、CGRP基本一致。由此认为四项指标的变化与脑血管损伤程度关系密切，其测定有助于判断病情。

Objective To evaluate the application of chemiluminescent microparticle immunoassay (CMIA) technology in quantitative detection of pm- transfusion infection indicators. Methods The pm- transfusion serum levels of hepatitis B surface antigen (HBsAg)，hepatitis C virus antibody (anti-HCV)，anti- Treponema pallidum- specific antibody (Anti-TP) and anti - humanimmunodeficiency virus antibody ( anti - HIV ) in 22，235 cases of patients were determined with chemiluminescent microparticle immunoassay ( CMIA) . The qualitative detection of all positive samples were carried out by the corresponding enzyme- linked immunosorbent assay (ELISA)，and the anti - treponema pallidum antibody ( Anti-TP) positive samples were confirmed by Treponema pallidum antibodies gelatin particle agglutination test (TPPA) . Results The number of positive cases of HB- sAg, Anti - HCV，Anti-TP and Anti- HIV in 22 ,235 cases of patients were 3028，336，684 and 63 respectively. The qualitative positive cases among these positive samples detected by ELISA were 2472, 269, 668, and 58. 603 samples in 684 cases of Anti-TP the quantitative positive samples were confirmed as positive by TPPA. Conclusion CMIA assay for quantification of pre-transfusion infection indicators could be used as early detection of infection “window period” to reduce the missing rate. It would be very helpful in the early diagnosis and treatment of patients, and self- protection of healthcare workers.

To determine changes in serum concentrations of total and free testosterone in young and middleaged men. the healthy men (n=126) were divided 2029yr, 3039yr and 4049yr three groups, their serum levels of total testosterone (T), free testosterone (FT), luteinizing hormone (LH), folliclestimulating hormone (FSH), sex hormone binding globulin (SHBG) and estradiol (E₂) were detected. The results were statically analyzed. The results showed that the levels of serum T and FT was found significantly decreased in 3039yr group than in 2029yr group (15.06±3.13 nmol/L vs 20.41±6.86 nmol/L, P＜0.01; 9.36±2.95 pg/L vs 11.48±3.88 pg/L, P＜0.05; respectively). There were young trends that agerelated decline in androgen levels.

Abstract: Objective To establish a quantitative detection method for determination of serum carcinoembryonicantigen levels. Methods Based on the principle of double antibody sandwich method, the acridinium ester chemiluminescence immunoassay was developed by using bridged avidin-biotin technique. Results The lowest detection limit of assay was 0.2 ng/mL. The linear range was 2.2～360 ng/mL and batch variation was 1.9%～2.2%. The total variance was 6.5%～8.1%. The test results by this assay and ROCHE varcinoembryonic antigen assay Kit were linear correlation (Y=0.9969X+0.6168, r=0.9921). Conclusion The assay developed in this study might meet the requirements of clinical test and are suitable for clinical application.

Objective To establish a subcutaneous xenograft model of human prostate carcinoma in nude mice. Methods PC-3 cells of the human prostate carcinoma were inoculated subcutaneously into the neck back of BAlB/c nude mice. The tumor incubation period, forming tumor percentage was calculated and pathological identification of tumors were carried out Results The mean incubation period of xenograft by inoculating subcutaneously with human prostate PC-3 cells was 24 days with 100% tumor forming. The time of tumor volume doubled was 10 days. Morphological and functional features of xenograft were basically same as original tumor. Conclusion The xenograft model of human prostate carcinoma established in this study could provide valuable experiment platform for radioimmunoimaging and radioimmunotherapy of the prostate carcinoma.

To evaluate the value of human epididymis secretory protein 4 (HE4) and CA125 in monitoring recurrence and prognosis of ovarian cancer. All individuals enrolled in the test must be above 60. The serum HE4 and CA125 levels were determined in pathology proved ovarian cancer group (75 cases), benign ovarian diseases group (73 cases), other malignant tumor group (53 cases) and healthy women group (103 cases). Follow-up studies were performed in 12 patients with ovarian cancer and the serum levels of HE4 and CA125 were detected. The results showed that the specificity of HE4 in the detection of elder ovarian cancer was higher than that of CA125. The serum HE4 level was decreased as the remission of the disease, and it decreased more quickly than that of serum CA125. In case of recurrence of the disease, the expression of serum CA125 delayed up to 3-6 months than that of HE4. The poorer prognosis was occurred in patients with a high serum HE4 level. The specificity of HE4 in the detection of ovarian cancer in elder women is satisfied and it may be acted as a good indicator for the remission, recurrence and prognosis of ovarian cancer.

 Lung cancer is the most common cause of cancer death worldwide. Early diagnosis and early treatment are of great importance. Lung cancer is a multi-factorial disease, and multiple genes involved in the development of complex multi-stage process. In recent years, domestic and foreign scholars on the early diagnosis of lung cancer biomarkers have done a lot of useful exploration. In this paper, recent developments in this field are reviewed.

近来年研究表明,尿微量白蛋白不仅是肾脏血管内皮损伤的标志,也是全身血管内皮细胞损伤的标志。一些调查研究认为,尿微量白蛋白阳性是心血管事件发生的独立危险因素^[1]。因此,检测尿微量白蛋白对相关疾病的早期诊断和治疗有重要的参考意义。为了验证免疫透射比浊法(简称“透射法”)微量白蛋白诊断试剂用于尿微量白蛋白检测的准确性及实用性,本文以免疫散射比浊法(简称“散射法”)微量白蛋白诊断试剂作参比方法,对检测结果进行相关性分析和方法学评价。

The incidence and mortality rate of the primary liver cancer are very high, and its postoperative metastasis and recurrence have become important factors that affect the prognosis of patients. Clinically, the detection methods to effectively predict and prevent of the liver cancer metastasis are in urgent need. Due to their advantage of real-time and noninvasive monitoring, circulating tumor cells (CTC), as a new tumor marker, play an important role in the early diagnosis, early treatment and individualized treatment, which induce the interest of the researchers. It has been recognized in clinical due to the successful applications in breast cancer, prostate cancer, colorectal cancer, lung cancer and other diseases. The latest research shows that the state of a liver cancer patient and the number of CTCs in blood are closely related. However, the diverse detection methods and their sensitivity, specificity limit their clinical applications. At present, the popular detection system mainly contains two aspects: the enrichment and identification. In recent years, new CTC detection methods appear and are used in clinical research. This article will review the related research.

To investigate the proportion of increasing carcinoma-embryonic antigen(CEA)in patients with malignant and benign diseases, in order to explore its auxiliary diagnosis in clinic, the data of 557 increasing CEA patients in hospital from January to May in 2010 were analyzed retrospectively with SPSS. The results showed that the patients with tumor were 77.74% and the non-tumor patients were 22.26% in all 557 increasing CEA samples. The values of CEA increased had significantly statistics difference between two groups (P＜0.01).In the CEA concentration range of 5.01~10.00μg/L, there is no significant difference between malignant and benign diseases groups (P＞0.05). The patients with hepatic carcinoma was different significantly from the patients with gastric cancer, lung cancer and colorectal cancer groups respectively (P＜0.01). The patients with breast cancer was also different from that of patients with hepatic carcinoma group (P＜0.05). The CEA was not a specific tumor marks. It could be also increased in non tumor patients in the CEA concentration range of 5.01~10.00μg/L. When the CEA might concentration is more than 10.00μg/L, it might be more useful in the diagnosis of malignant and benign diseases.

Objective To evaluate the method for thyrotropin receptor antibody（TRAb）measurement with Roche Electro chemiluminescence (ECL) system and to establish the reference range in our laboratory. Methods The within-run and between-day imprecision was performed with high and low lever mixed serum. The analytic measure range (AMR) was evaluated with serially diluted serums. Limit of quantity (LoQ) was evaluated with continuous detection of low level serums. 20 normal sera were random taken to verify the reference range. 50 normal human serum and 50 patient’s serum samples were used to verify the specificity and sensitivity of method. Results The imprecision of within-run for high and low concentration of TRAb sample were 0.63% and 6.13%. The imprecision of between-day for high and low value concentration of TRAb sample were 4.15% and 9.55%. Reportable low limit was 1.20IU/L. The analytical measurement range (AMR) was 2.72-33.66 IU/L. the reference range was less than 1.24 IU/L. The specificity and sensitivity of measurement were 94% and 92% respectively. The bias of two detection system measurement data was less than the acceptable biases of the laboratory. Conclusion The Electro chemiluminescence system for the TRAb measurement meets the requirement of CAP.

The signal pathway of apoptosis induced by nuclear DNA damage is getting explicit gradually. But the relationship between the mitochondrial DNA damage and apoptosis remains unclear. The rate of apoptosis is influenced significantly by the break, depletion and dysfunction of mitochondrial DNA. The reaction of ρ⁰ cell to several of apoptotic inducible factors is different from its parental cell lines. The generation of ROS is not the essential condition of the mitochondrial DNA damage induced apoptosis and the cascade may be initiated by mitochondrial DNA damage itself. In a word, further study will be needed to know what role does the mitochondrial DNA damage act in the signal pathway.

To investigate the clinical value of combination detection serum tumor markers SCC, CA125, CA15-3 and CA19-9 in the diagnosis of cervical cancer, the serum SCC, CA125, CA15-3 and CA19-9 levels in 31 pretreatment cervical cancer patients, 22 patients with cervical benign diseases and 53 healthy subjects were detected. The results showed that the serum SCC, CA125 and CA15-3 levels in cervical cancer patients were significantly higher than in patients with cervical benign diseases and healthy control (P＜0.05). The serum SCC among four serum tumor markers used alone in the diagnosis of cervical cancer showed a highest sensitivity (41.90%). Combination detection of SCC and CA125, the sensitivity was increased to 58.1%, the specificity was 72.0%. Combination detection of serum SCC and CA125 would improve the sensitivity in cervical cancer detection. It would be helpful for the assistant diagnosis of cervical cancer.

Objective To investigate the radiolabelling method of ⁹⁹Tc^m-mAb109 against breast cancer, and evaluate its biodistribution and imaging property on tumor-bearing animal model. Methods Monoclonal antibody mAb109 was reduced by 2-mercaptoethanol, the concentration of reduced antibody and the number of sulfhydroxyl group on reduced antibody were determined. The radiolabelling of mAb109 with ⁹⁹Tc^m was carried out by using transchelating method through glucohepatonate. The labeling yield and stability of radiolabelled mAb109 were measured. The biodistribution and imaging study of ⁹⁹Tc^m-mAb109 were carried out on a breast tumor-bearing animal model. Results The reduced antibody mAb109 remained high integrity. The average of 5.38 sulfhydroxyl groups per reduced antibody was detected. The radiolabelling yield of ⁹⁹Tc^m-mAb190 was more than 90%, and ⁹⁹Tc^m-mAb190 was very stable in vitro. Radiolabelled antibody significantly accumulated on tumor sites and remained for long time. Tumor-bearing animal model showed clear imaging after injection of ⁹⁹Tc^m-mAb109. Conclusion The radiolabelling method of ⁹⁹Tc^m-mAb109 is successfully established, and ⁹⁹Tc^m-mAb109 shows potentially targeting characters for animal model borne glucose-related protein 78 positive breast tumors and is worthy for further study.

为比较化学发光免疫分析（CLIA）、放射免疫分析（RIA）和磁珠酶免疫分析（MSP-ELISA）测定血清TT3、TT4、FT3、FT4 和TSH的方法学特性，分别用CLIA、RIA和MSP-ELISA测定标准品、质控品及40份患者血清中TT3、 TT4、FT3、FT4 和TSH含量，进行线性、相关性、精密度、回收率等实验结果分析评价。结果显示：CLIA、RIA和MSP-ELISA检测TT3、TT4、FT3、FT4和TSH的线性和相关性无显著性差异，CLIA检测的精密度、回收率优于RIA、MSP-ELISA，且具有快速的特点。

1971年瑞典斯德哥尔摩大学的Engvall和Perlmann ^[1]首次建立了酶联免疫吸附测定( Enzyme-Linked Immunosorbnent Assay,ELISA)方法,用于定量检测家兔血液中的IgG。四十年来,随着单克隆技术的发展应用及免疫试剂盒的商品化,ELISA已广泛应用于临床医学、动物检疫、食品科学、植物病毒、药物残留等分析领域,这一技术的普遍应用归功于商品试剂盒和自动或半自动检测仪器的研发与问世^[2]。ELISA不仅在临床检验工作中占有举足轻重的地位,它在医学基础研究中的应用也越来越普遍并发挥着重要作用。目前关于ELISA原理及方法应用的文章已经很多,其中不乏内容完备者,但是,我们在承担心血管领域科研课题进行ELISA检测过程中又发现了很多的基本问题,例如:许多研究者因为对ELISA了解不够全面深入,在收集样本时处理不当、购买何种检测试剂盒、如何计算所需试剂盒的数量以及在ELISA检测过程中需要注意的各种问题,等等。上述问题看似琐屑,实际操作不当却会直接影响测定结果的准确性。本文旨在总结ELISA操作的实际经验,从基础研究的角度出发,简要介绍ELISA实验方法的基本原理、分类及应用,重点讲述在基础医学研究中应用ELISA方法时需要注意的一些事项,以便为研究者提供一定的参考和帮助。

Objective To investigate the distribution feature of dystrophin gene deletion mutations in Chinese Han Duchenne/Becker muscular dystrophy (DMD/BMD)pediatric patients. Methods Determination of deletion mutation in dystophin gene of 964 DMD/BMD suspected pediatric patients was performed by amplifying 20 segments of major deletion “hot spot” region with polymerase chain reaction (PCR).The deletion pattern of dystophin gene was consequently summarized and the distribution of breakpoints of dystophin gene deletion mutation was also analyzed.Results The total 491cases (51.0%) had confirmed deletion mutations. Mutations in 75 cases (15.2%) located in the region of 5’-flanking region, 402 cases (81.7%) in the region of central region,and 13 cases (2.6%) deletions span in both 5’-flanking and central regions.The most common deletions including exon 50, 49 and 48 deletions ranked the top. 74％of deletion breakpoints fell in introns 44~50 in dystophin gene, in which about 21% of deletion breakpoints fell in intron 44. Conclusion The study indicates that more than half of suspected DMD/BMD cases are confirmed deletion mutation in dystophin gene. Deletion mutations are mostly distributed in the central region of DMD gene in Chinese Han pediatric population.

To determine the preferred and effective CD4 antibody used for T helper (Th) cell study, Th cells before and after stimulation were investigated by flow cytometry with three different fluorochrome-labeled CD4 antibodies. Before and after stimulation with PMA and Ion, the percentage of Th cells (CD3⁺CD8^- T cells) was determined by CD3-APC and CD8-PerCP, while the percentage of Th cells (CD4⁺ T cells) was detected by CD4-PerCP, CD4-FITC and CD4-APC. Furthermore, CD4⁺ T cells sorted by immunomagnetic beads were used to confirm the efficacy of CD4-APC antibody in Th cell research. The results showed that there was no significant difference in the percentages of Th cells (CD3⁺CD8^- T cells) before and after stimulation (P＞0.05). The percentages of Th cells (CD4⁺CD8^- T cells) before and after stimulation detected by three fluorochrome-labeled CD4 antibodies were all declined. Among them, CD4-PerCP declined significantly (P＜0.05), while CD4-FITC and CD4-APC declined slightly (P＞0.05). The MFI of CD4-APC was the strongest among three fluorochrome-labeled CD4 antibodies and did not decrease significantly after stimulation (P＞0.05), while the MFI of CD4-PerCP and CD4-FITC decreased sharply (P＜0.05). Cells sorted by immunomagnetic beads also confirmed that the stimulation had little effect on the expression of CD4-APC. It is thus showed that the preferred CD4-APC antibody was effective in Th cell research among the three fluorochrome-labeled CD4 antibodies.

Objective To establish the normal reference range of serum tumor markets for health people in Shanghai to provide the basis for the clinical application. Method 604 healthy human serum which were collected from March 2012 to April 2013 have been tested by Flow Fluorescence Immunoassay for the concentration of AFP, CA125, CYFRA21-1, CEA, NSE, and CA19-9. Results The AFP reference range was <10.32ng/mL in male and <8.12ng/mLin female; CA125 reference range was 27.02U/mL; CEA reference range was <4.3ng/mL in male and <3.74ng/mLin female; NSE reference range was <11.64ng/mL in female with age less than 45years, <15.20ng/mL in female with age more than 45years, <18.47ng/mL in male wit age less than 45years and <20.04ng/mL with age more than 45 years; CA19-9 reference range was < 20.73U/mL; and tPSA reference range was <1.48ng/mL.Conclusion The preliminarily reference range of AFP, CA125, CYFRA21-1, CEA, NSE, CA19-9 for healthy people in Shanghai are established.

Objective To establish a stable and effective system of mouse embryonic fibroblasts (MEFs) feeder layer in order to culture mouse embryonic stem cells(ESC)in vitro.Methods Embryonic fibroblast cells for primary culture were derived from mouse embryos (pregnant 13-15 days) through trypsin digestion. Results After being treated with mitomycin C (10μg/mL), MEFs proliferation could be efficiently repressed and be made into the feeder layer for ESC Undifferentiated growth. Conclusion MEFs feeder layer could effectively support mouse embryonic stem cells to grow.

Objective By analyzing 417 high risk hs-CRP health examination indicators in our hospital, to understand the incidence of different diseases and to explore the reason and preventive measures. Methods Retrospective analysis was used to analyze the hs-CRP. Based on the high risk hs-CRP indicator, six kinds of diseases including hypertension, diabetes, carotid atherosclerosis, cerebral infarction, coronary heart disease and fatty liver disease and the gender were selected for statistical analysis. Results The results of body mass index in the inflammation group showed significant differences between male and female group. The high risk hs-CRP of fatty liver disease and hypertension was particularly severe. In addition to the carotid atherosclerosis group, the remaining five kinds of diseases showed significant difference between men and women group. Conclusion The abnormal increase of hs-CRP may due to many reasons; the presence of other disease factors should be took into account in clinical testing.

Objective To explore the clinical relevance of combined detection of thin prep liquid-based cytology (TCT) and human papillomavirus (HPV) DNA in screening for early cervical lesions. Methods The thin prep liquid-based cytology detection was carried out in 375 cases of HPV-DNA positive patients. The results of TCT and HPV-DNA were analyzed by TBS reporting method. Results The number of benign reactive changes was 273 (72.8%) in TCT examiner of 375 cases of HPV-DNA positive patients. The number of ASC positive was 84 cases (22.4%), LSIL positive was 6 cases (1.6%) and HSIL-positive was 12 cases (3.2%). The infection rates of sub-type of HPV high-risk were 69.23%, 92.86%, 100% and 100% respectively. Conclusion The combined detection of TCT with HPV-DNA could improve the diagnosis rate for early cervical lesions.

Abstract: After decades of development, radionuclide therapy,as a primary part of nuclear medicine,has become one of the important clinic treatments for some special diseases. Recently,radionuclide therapy has been paid more and more attention. The development of radionuclide therapy and the commonly used treatments are reviewed in this paper.

Objective To analyze the related effect factors on the serum pepsinogen (PG) levels in healthy people checkup, and to explore the management measure for the people with abnormal PG. Method 531 people who took physical examination were involved in this study. The serum PG levels were detected by time resolved f1uoroimmunoassay. The rate of HP infection was detected by ¹³C urea breath test. Results The serum PGⅠ level in males was significantly higher than that of females, while the detected rate of women with PGⅠ<60μg/L and the PG/PGⅡ <6 was significantly higher than that of men. In the age of 20~70 group, the serum PG I and PGⅡ levels were increase gradually as the age increase, while the PGⅠ/ PGⅡ was decreased. The serum PGⅠand PGⅡ levels in HP positive group were significantly higher than that of HP negative group, while the PG/PGⅡ levels were significantly lower than that of HPnegative group. Conclusion The serum PG levels were related with age, gender and HP infection. It is of great importance and significance to combine the determination of serum PGⅠ, PGⅡ and PGⅠ/ PGⅡ ratio for scanning stomach disease in healthy people.

Objective To investigate the value of detection of urinary catecholamine metabolites, serum carcinoembryonic antigen (CEA) and neural enolase (NSE) in the diagnosis and differential diagnosis of neuroblastoma (NB). Method The urinary catecholamine metabolites including vanillylmandelic acid (VMA) and homovanillic acid (HVA), and serum CEA and NSE levels in 36 neuroblastoma patients and 30 controls were detected by high-performance liquid chromatography and chemiluminescence assay respectively. Results The urinary catecholamine metabolites VMA, HVA, serum CEA and NSE in NB patients were significant higher than those in the control group (P<0.01). The positive rate of VMA and HVA in NB patients was 86.1% and 75.0 % respectively. The positive rate of combination detection of VMA and HVA was increased to 91.7%. The combined detection with serum NSE or CEA, the positive rate increased to 94.4%, while combined detection with both serum NSE and CEA, the positive was increased to 97.2%. Conclusion The detection of urinary catecholamine metabolites VMA, HVA, serum CEA and NSE level play an important role in the diagnosis and differential diagnosis of neuroblastoma. The combination detection could improve the positive rate.