The Oncogenic Role of KLF7 in Colon Adenocarcinoma and Therapeutic Perspectives.

Colon adenocarcinoma, a highly prevalent and aggressive form of colorectal cancer, necessitates a comprehensive understanding of its molecular mechanisms to identify potential therapeutic targets. The Krüppel-like factor 7 (KLF7), a transcription factor, has been associated with various malignancies, yet its specific role in colon adenocarcinoma remains largely unexplored. Here, we aimed to determine the expression and functional significance of KLF7 in colon adenocarcinoma. Our findings revealed a significant upregulation of KLF7 expression in colon adenocarcinoma tissues compared to adjacent normal tissues. Moreover, elevated KLF7 expression correlated with advanced tumor stage, lymph node metastasis, and poor overall survival in colon adenocarcinoma patients. Functional assays demonstrated that silencing KLF7 resulted in reduced cell proliferation, migration, and invasion, indicating its involvement in promoting tumor growth and metastasis. Additionally, we identified potential downstream targets of KLF7, including genes associated with cell cycle regulation and epithelial-mesenchymal transition. These results underscore the tumor-promoting role of KLF7 in colon adenocarcinoma, positioning it as a potential prognostic biomarker and therapeutic target for this aggressive disease.

Effect of different surgical approaches on the survival and safety of Siewert type II esophagogastric junction adenocarcinoma: a systematic review and meta-analysis.

BACKGROUND: Whether a transthoracic (TT) procedure by a thoracic surgeon or a transabdominal (TA) by a gastrointestinal surgeon is best for Siewert type II esophagogastric junction adenocarcinoma (EGJA) remains unknown. Survival and perioperative outcomes were compared between the two groups in this meta-analysis to clarify this argument. METHODS: We searched 7 databases for eligible studies comparing TT and TA procedures for Siewert type II EGJA. The final analyzed endpoints included intraoperative and hospitalization outcomes, recurrence, complication, and survival. RESULTS: Seventeen studies involving 10,756 patients met the inclusion criteria. The TA group had higher rates of overall survival (OS) (HR: 1.31 [1.20 ~ 1.44], p < 0.00001) and disease-free survival (DFS) (HR: 1.49 [1.24 ~ 1.79], p < 0.0001). The survival advantage of OSR and DFSR increased with time. Subgroup analysis of OS and DFS suggested that TA remained the preferred approach among all subgroups. More total/positive lymph nodes were retrieved, and fewer lymph node recurrences were found in the TA group. The analysis of perioperative outcomes revealed that the TA procedure was longer, had more intraoperative blood loss, and prolonged hospital stay. Similar R0 resection rates, as well as total recurrence, local recurrence, liver recurrence, peritoneal recurrence, lung recurrence, anastomosis recurrence and multiple recurrence rates, were found between the two groups. The safety analysis showed that the TT procedure led to more total complications, anastomotic leakages, cases of pneumonia, and cases of pleural effusion. CONCLUSIONS: The TA procedure appeared to be a suitable choice for patients with Siewert type II EGJA because of its association with longer survival, fewer recurrences, and better safety.

The antimicrobial peptide chensinin-1b alleviates the inflammatory response by targeting the TLR4/NF-κB signaling pathway and inhibits Pseudomonas aeruginosa infection and LPS-mediated sepsis.

Excessive inflammatory responses are a major contributor to the high mortality associated with sepsis, a prevalent global complication. Therefore, the potential therapeutic strategy for sepsis involves targeting macrophages and reducing proinflammatory cytokine release. Chensinin-1b, an analog of the natural antimicrobial peptide derived from Rana chensinensis skin secretion, exhibits broad-spectrum antibacterial activity and adopts a random coil conformation in both PBS and membrane solution. By efficiently neutralizing LPS, chensinin-1b holds promise in alleviating LPS-induced inflammatory responses. In this study, we established a mouse septic shock model by exposing mice to multiple-drug-resistant Pseudomonas aeruginosa, as well as an endotoxin-mediated sepsis model induced by LPS. Administering chensinin-1b significantly prolonged the survival of the experimental mice, concurrently mitigating inflammatory responses and reducing organ damage. Additionally, we investigated the anti-inflammatory mechanism of chensinin-1b using a constructed LPS-induced mouse macrophage RAW264.7 inflammatory model. Our findings demonstrated that chensinin-1b effectively mitigated the excessive activation of the TLR4/NF-κB signaling pathway by directly neutralizing extracellular LPS, thus ameliorating the inflammatory response. Moreover, upon blocking the TLR4 signaling pathway, chensinin-1b further reduced the release of proinflammatory cytokines induced by LPS, indicating alternative modes of regulation. Notably, chensinin-1b rapidly entered RAW264.7 cells within 30 min via endocytosis, diffusing into the cytoplasm while retaining its anti-inflammatory properties intracellularly. Although further investigations are warranted to comprehensively elucidate the intracellular anti-inflammatory mechanism of chensinin-1b, our findings substantiate its possession of anti-inflammatory properties both intracellularly and extracellularly. Thus, chensinin-1b emerges as a promising candidate for mitigating excessive inflammatory responses associated with sepsis.

Risk prediction of intraoperative pain in percutaneous microwave ablation of lung tumors under CT guidance.

OBJECTIVES: To evaluate the effect of intraoperative pain in microwave ablation of lung tumors (MWALT) on local efficacy and establish the pain risk prediction model. METHODS: It was a retrospectively study. Consecutive patients with MWALT from September 2017 to December 2020 were divided into mild and severe pain groups. Local efficacy was evaluated by comparing technical success, technical effectiveness, and local progression-free survival (LPFS) in two groups. All cases were randomly allocated into training and validation cohorts at a ratio of 7:3. A nomogram model was established using predictors identified by logistics regression in training dataset. The calibration curves, C-statistic, and decision curve analysis (DCA) were used to evaluate the accuracy, ability, and clinical value of the nomogram. RESULTS: A total of 263 patients (mild pain group: n = 126; severe pain group: n = 137) were included in the study. Technical success rate and technical effectiveness rate were 100% and 99.2% in the mild pain group and 98.5% and 97.8% in the severe pain group. LPFS rates at 12 and 24 months were 97.6% and 87.6% in the mild pain group and 91.9% and 79.3% in the severe pain group (p = 0.034; HR: 1.90). The nomogram was established based on three predictors: depth of nodule, puncture depth, and multi-antenna. The prediction ability and accuracy were verified by C-statistic and calibration curve. DCA curve suggested the proposed prediction model was clinically useful. CONCLUSIONS: Severe intraoperative pain in MWALT reduced the local efficacy. An established prediction model could accurately predict severe pain and assist physicians in choosing a suitable anesthesia type. CLINICAL RELEVANCE STATEMENT: This study firstly provides a prediction model for the risk of severe intraoperative pain in MWALT. Physicians can choose a suitable anesthesia type based on pain risk, in order to improve patients' tolerance as well as local efficacy of MWALT. KEY POINTS: • The severe intraoperative pain in MWALT reduced the local efficacy. • Predictors of severe intraoperative pain in MWALT were the depth of nodule, puncture depth, and multi-antenna. • The prediction model established in this study can accurately predict the risk of severe pain in MWALT and assist physicians in choosing a suitable anesthesia type.

Preoperative computed tomographic-guided percutaneous coil localization of pulmonary nodules: 184 Cases in Single-center.

OBJECTIVES: The aim of this study was to evaluate the feasibility, accuracy, and safety of computed tomographic (CT)-guided coil localization before video-assisted thoracoscopic surgery. METHODS: One hundred and eighty-four consecutive patients with 200 pulmonary nodules who underwent CT-guided percutaneous coil localization before thoracoscopic surgery were retrospectively analyzed in this study. Success rate for localization, complication rates, CT findings, and pathological results of the lesions, as well as the data related to surgery were all recorded and analyzed. RESULTS: All 184 patients with 200 lesions completed localization and resection. The success rate of the coil localization on lesion baseline was 99.0% (198/200) and 98.9% (182/184) on patient baseline. The number of wedge resection, segmental resection, and lobectomy were 179 (89.5%), 19 (9.5%), and 2 (1.0%), respectively. The malignancy rates in a lesion-based analysis were 83.5% (91.1% in ground-glass nodules, 91.4% in part-solid nodules, and 37.9% in solid nodules). No serious complications occurred in all localization procedures. CONCLUSION: Preoperative CT-guided percutaneous coil localization is a safe and effective method to facilitate high success rates for both wedge and segmental resection for peripheral pulmonary nodules.

Laparoscopic versus open radical resection for transverse colon cancer: evidence from multi-center databases.

BACKGROUND: The role of laparoscopic approach is still a controversy for transverse colon cancer. Our investigation aimed to evaluate the perioperative and oncologic outcomes of laparoscopic versus open radical resection for transverse colon cancer based on evidence from multi-center databases. METHODS: 416 patients with transverse colon cancer undergoing radical surgery were analyzed including 181 laparoscopic resections and 235 open resections from January 2004 to May 2017 based on multi-center databases. Perioperative and oncologic outcomes were compared. RESULTS: No statistical differences regarding the baseline characteristics were observed between the two groups except the procedure type. Compared with open approach, laparoscopic approach was associated with statistically longer operation time (209.96 vs. 173.31 min, P = 0.002), significantly shorter time to soft food intake (4.73 vs. 6.01 days, P = 0.034), and shorter postoperative hospitalization (12.05 vs. 14.44 days, P = 0.001). In terms of oncologic outcomes, laparoscopic resection was correlated with statistically more lymph node retrieval (13.52 vs. 15.91, P = 0.002) and similar 5-year overall survival (91.2% vs. 89.1%, P = 0.356) and disease-free survival (89.6% vs. 86.0%, P = 0.873), compared with open resection. CONCLUSIONS: For patients with transverse colon cancer, laparoscopic approach can achieve several short-term advantages without decreasing long-term oncologic survival.

Predictive value of peripheral blood NLR and PLR in perioperative period on the prognosis of patients with intrahepatic cholangiocarcinoma / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨肝内胆管癌（intrahepatic cholangiocarcinoma，ICCA）围手术期外周血中性粒细胞与淋巴细胞比率（neutrophil-to-lymphocyte ratio，NLR）和血小板与淋巴细胞比率（platelet-to-lymphocyte ratio，PLR）对患者预后的预测价值。方法：收集2015年1月至2018年1月在上海市松江区中心医院接受肝切除术治疗的ICCA患者97例作为ICCA组，选择同期在本院做健康体检的志愿者100例作为正常对照组。检测两组受试者术前1 d、术后3 d和7 d外周血的NLR、PLR，采用单因素、多因素分析ICCA患者术后随访期死亡的危险因素，采用Kaplan-Meier生存曲线分析术后3 d的NLR和PLR对ICCA患者术后生存时间的影响，采用受试者工作特征曲线（receiver operating characteristic curve，ROC）分析术后3 d的NLR和PLR水平对患者术后随访期死亡的预测价值。结果：ICCA组患者术前1 d、术后3和7 d外周血的NLR、PLR均高于正常对照组（均P<0.05），术前1 d和7 d外周血NLR、PLR差异无统计学意义（P>0.05），术后3 d外周血NLR、PLR水平最高（P<0.05）。多发肿瘤、合并淋巴结转移、TNM分期Ⅲ~Ⅳ、CA199水平增高、术后3 d的NLR和PLR较高分别是ICCA患者随访期死亡的独立危险因素（均P<0.05）。ROC曲线显示，术后3 d的NLR和PLR高低对ICCA患者术后生存时间具有预测价值。Kaplan-Meier生存曲线显示，低NLR[（50.32±3.69） vs （30.12±2.36）个月]和低PLR[（53.6±3.75） vs （37.6±2.96）个月]患者生存时间均长于高NLR和PLR的ICCA患者（均P<0.05）。结论：ICCA术后3 d的NLR和PLR异常增高是患者肝切除术后死亡的独立危险因素，其对患者生存时间具有早期预测价值。

Capn4 promotes colorectal cancer cell proliferation by increasing MAPK7 through activation of the Wnt/ß-Catenin pathway.

Increasing evidence has suggested that Capn4 is upregulated and functions as a potential tumor promoter in several human cancer types. However, the potential biological roles and regulatory mechanisms of Capn4 in colorectal cancer (CRC) remains unclear. Here, we found that Capn4 expression was elevated in CRC tissues than adjacent noncancerous tissues. Additionally, we also found that overexpression of Capn4 is significantly correlated with tumor progression and poor survival in CRC patients. Furthermore, our experimental data revealed that increased expression of Capn4 was observed in CRC cell lines and ectopic expression of Capn4 significantly enhanced in vitro cell proliferation, whereas knockdown of Capn4 suppressed CRC cells growth in vitro and in vivo. Moreover, our results indicate that Capn4 promotes cell proliferation by increasing MAPK7 expression, which has been reported to control the proliferation of many cancers. Mechanistically, Capn4 upregulates MAPK7 expression through activation of the Wnt/ß-Catenin pathway in CRC cells. Therefore, we identified a tumorigenic role of Capn4 in CRC and suggested a potential therapeutic target for CRC patients.

Expression and clinical significance of Flotillin-2 in gastric cancer tissues / 中国肿瘤生物治疗杂志

@# Objective: To investigate the expression of Flotillin-2 (Flot-2) protein in gastric cancer tissues and its relationship with clinicopathological features and prognosis of gastric cancer (GC) patients. Methods: 112 samples of gastric cancer tissue and the corresponding paracancerous tissue that resected at the gastrointestinal surgery department of the Second Affiliated Hospital of Nanchang University between January 2009 andApril 2010 were collected for this study. The expression of Flot-2 protein in tumor tissues was detected by immunohistochemistry. The survival data were analyzed by Kaplan-Meier and Log-Rank test, and the survival curve was plotted. Spearman correlation analysis was used to examine the relationship between Flot-2 protein expression and clinicopathological characteristics and prognosis of GC patients. Results: In gastric cancer tissues, Flot-2 was primary stained in cytoplasm. Level of Flot-2 was significantly higher in gastric cancer tissues compared with that in paracancerous tissues (53.57% vs 46.43%, P<0.05). Expression of Flot-2 in tumor tissues was significantly associated with tumor size, depth of invasion, lymph node metastasis, distant metastasis and AJCC stage (all P<0.01), but not with gender, age, differentiation degree and tumor location (P>0.05). Moreover, survival analysis showed that the overall survival of patients with low Flot-2 expression was significantly higher than that of the patients with high level (P<0.01). Cox regression analysis indicated that distant metastasis, AJCC stage and Flot-2 expression were the independent risk factors for the prognosis of GC patients. Conclusion: Flot-2 protein was highly expressed in gastric cancer tissues and closely correlated with the poor prognosis of GC patients; Flot-2 is an independent risk factor for GC prognosis and may be served as a potential therapeutic target for gastric cancer.

Adaptive Energy-Efficient Target Detection Based on Mobile Wireless Sensor Networks.

Target detection is a widely used application for area surveillance, elder care, and fire alarms; its purpose is to find a particular object or event in a region of interest. Usually, fixed observing stations or static sensor nodes are arranged uniformly in the field. However, each part of the field has a different probability of being intruded upon; if an object suddenly enters an area with few guardian devices, a loss of detection will occur, and the stations in the safe areas will waste their energy for a long time without any discovery. Thus, mobile wireless sensor networks may benefit from adaptation and pertinence in detection. Sensor nodes equipped with wheels are able to move towards the risk area via an adaptive learning procedure based on Bayesian networks. Furthermore, a clustering algorithm based on k-means++ and an energy control mechanism is used to reduce the energy consumption of nodes. The extended Kalman filter and a voting data fusion method are employed to raise the localization accuracy of the target. The simulation and experimental results indicate that this new system with adaptive energy-efficient methods is able to achieve better performance than the traditional ones.

Chinese expert consensus workshop report: Guidelines for thermal ablation of primary and metastatic lung tumors.

Although surgical resection is the primary means of curing both primary and metastatic lung cancers, about 80% of lung cancers cannot be removed by surgery. As most patients with unresectable lung cancer receive only limited benefits from traditional radiotherapy and chemotherapy, many new local treatment methods have emerged, including local ablation therapy. The Minimally Invasive and Comprehensive Treatment of Lung Cancer Branch, Professional Committee of Minimally Invasive Treatment of Cancer of the Chinese Anti-Cancer Association has organized multidisciplinary experts to develop guidelines for this treatment modality. These guidelines aim at standardizing thermal ablation procedures and criteria for selecting treatment candidates and assessing outcomes; and for preventing and managing post-ablation complications.

Development of a competitive radioimmunoassay for glypican-3 and the clinical application in diagnosis of hepatocellular carcinoma.

OBJECTIVE: Glypican-3 (GPC3) is a promising specific tumor maker for hepatocellular carcinoma (HCC). The aim of this study is to establish a method to detect serum GPC3 and evaluate the clinical application on clinical diagnosis. DESIGN AND METHODS: A competitive radioimmunoassay for detecting serum GPC3 was developed. Clinical sera were detected by the proposed method and AFP, CA19-9 chemiluminescence immunoassay kit. RESULTS: The proposed method with high sensitivity, specificity and precision had no or little detectable cross-reactivity with relating tumor markers in the dynamic range from 15 to 500 ng/mL, and the detection limit was 0.5 ng/mL. The level of GPC3 in HCC was obviously higher than that in normal liver or other liver diseases. Additionally, our method showed high shows higher sensitivity and specificity for GPC3 than AFP and combined AFP/CA19-9. CONCLUSIONS: This paper provided an applicable competitive radioimmunoassay for GPC3 with high sensitivity, specificity and precision. In addition, using GPC3 for HCC diagnosis was more valuable than AFP.

Sequential treatment of superior vena cava syndrome caused by of non-small cell carcinoma lung cancer (NSCLC) with vascular stenting and iodine-125 implantation.

Feasibility and efficacy of sequentially performed endovascular stenting and Iodine-125 brachytherapy for malignant superior vena cava syndrome (SVCS) were evaluated. Thirty-four patients with malignant SVCS caused by NSCLC underwent sequential treatment of endovascular stenting and Iodine-125 brachytherapy. SVCS was diagnosed in all patients by CT images or vena-cavography. Pathology diagnosis was acquired by image guided biopsy. Endovascular stent placement was performed as first-line treatment for symptom relief. CT-guided Iodine-125 seed implantation performed 24hr after stenting. Clinical end points were resolution of symptoms and local efficacy of primary malignancy regression. Symptom relief rate was >90% after 24hr and 97% after 3 months. No migration of seeds or restenosis occurred in any patient. The local efficacy (defined as either partial or complete response) was 53%, 79%, 88% and 74% after 1, 3, 6 and 12 months, respectively. Mean SVCS-free survival time was 305 days (range 120-960 days). Two patients were still alive at the time of this writing, Thirty-one died from progression and one died from acute heart disease. Sequentially performed endovascular stenting and Iodine-125 brachytherapy provides a safe and effective alternative for malignant SVCS caused by NSCLC.

Development of a rapid and sensitive magnetic chemiluminescent enzyme immunoassay for detection of luteinizing hormone in human serum.

OBJECTIVES: A rapid and sensitive magnetic particles (MPs)-based chemiluminescence enzyme immunoassay (MPs-CLEIA) for determination of luteinizing hormone (LH) in human serum was developed and validated. DESIGN AND METHODS: The method was developed based on a sandwiched immunoreaction format, in which MPs served as both the solid phase and separator. The horseradish peroxidase(HRP)-luminol-H(2)O(2) chemiluminescent system was chosen as the detection system. RESULTS: This method had a detection limit of 0.2 mIU/mL and a linear range of 0.5-200 mIU/mL. The assay showed little cross-reactivity, and the intra- and inter-assay coefficients of variation were less than 10%. The recoveries were from 101.6% to 104.1%. The method has been successfully applied to detection of LH in human serum and showed good correlation compared with radioimmunoassay (RIA). CONCLUSION: The MPs-CLEIA provided apparent advantages over RIA, and facilitated the development of high-throughput screening and automated operation systems in clinical diagnosis.

Magnetic particle-based chemiluminescence enzyme immunoassay for free thyroxine in human serum.

A magnetic particles-based chemiluminescence enzyme immunoassay with high sensitivity, specificity, rapidity, and reproducibility was developed for the determination of free thyroxine in human serum. A competitive assay has been proposed with horseradish peroxidase labeled thyroxine analog. The immunomagnetic particles coated with anti-fluorescein isothiocyanate antibody was used as dispersed solid phase and separation means for the immunoassay. Experimental conditions, such as temperature, the volume of magnetic particles and substrate, incubation time, dilution ratio and other relevant variables upon the immunoassay have been examined and optimized. The proposed method exhibited high performance which the linear range was 1.59-122 pmol L(-1) and the detection limit was 0.25 pmol L(-1). A coefficient of variance of less than 15% was obtained for both intra-assay and inter-assay precision. The present method has been successfully applied to the analysis of free thyroxine in human serum. The diagnostic accordance rate of the method for normal serum, hyperthyroidism and hypothyroidism are satisfactory. Good correlations were obtained between the results by the proposed method and the commercial radioimmunoassay kit. The present method exhibits good potential in the fabrication of FT4 diagnostic kits which could be used in the clinical analysis and facilitated the development of automated operation systems in the clinical practice.

Development of magnetic particle-based chemiluminescence enzyme immunoassay for the detection of 17beta-estradiol in environmental water.

In the present work, a simple, fast, and highly sensitive chemiluminescence enzyme immunoassay for 17beta-estradiol (E2) in environmental water samples was developed, using magnetic particles (MPs) labeled with secondary antibody as both the immobilization matrix and the separation tools. The specific anti-E2 polyclonal antibody (PcAb) was produced against a conjugate of estradiol-bovine serum albumin. The specificity of the anti-E2 antibody was studied. The results showed that the antibody did not cross-react with the structurally related endocrine-disrupting compounds, including estrone, ethinyl E2, estriol, E2-17-glucuronide, E2-3-sulfate-17-glucuronide, androstenedione, and dihydrotestosterone. The water samples were pretreated with solid-phase extraction using C18 cartridges for the removal of matrix effects. Several physicochemical parameters including the dilution ratios of E2-6-horseradish peroxidase conjugate and anti-E2 PcAb, immunoreaction time, volume of chemiluminescent substrate and MPs, chemiluminescence reaction time, and pH of assay solution were studied and optimized. At optimal experimental conditions, it was found that the proposed method exhibited high performance with detection limit of 2.0 pg/mL, linear range of 20-1,200 pg/mL, and total assay time of 45 min. Both inter- and intra-assay coefficient of variation were less than 10%. The average recoveries of three different spiked concentration samples ranged from 86.3% to 108%. The method was successfully applied to the determination of E2 in river, waste, and tap water, and showed a good correlation with the commercially available radioimmunoassay kit.

Evaluation of alpha-fetoprotein (AFP) in human serum by chemiluminescence enzyme immunoassay with magnetic particles and coated tubes as solid phases.

In this work, the monoclonal antibodies (McAbs) to alpha-fetoprotein (AFP) were immobilized on two different solid phases, i.e., magnetic particles (MP) and coated tubes (CT). Based on this, a MP based chemiluminescence enzyme immunoassay (MP-CLEIA) and a CT based CLEIA (CT-CLEIA) were proposed for the evaluation of AFP in human serum and their analytical merits were studied and compared. By detailed discussion of several performance variants, including the concentration of immobilized McAb, dilution ratio of horseradish peroxidase (HRP) labeled McAb (HRP-McAb), total assay time, substrate volume, chemiluminescent kinetics, and hook effect concentration, the advantages of MP-CLEIA became conspicuously apparent. Moreover, in the presence of MP, the catalytic activity of labeled enzyme was kept to high extent and the stability of immunoreagents was satisfied. Finally, 59 human serum samples were detected by the MP-CLEIA and a good correlation was obtained when comparing the results with that from a commercial electrochemiluminescence immunoassay kit.

Development of a sensitive, rapid, biotin-streptavidin based chemiluminescent enzyme immunoassay for human thyroid stimulating hormone.

A highly sensitive "two-site" chemiluminescent immunoassay specific for human thyroid stimulating hormone (TSH) was developed. The signal amplification was achieved via a biotin-streptavidin system (BSAS). The HRP-luminol-H(2)O(2) chemiluminescent system with high sensitivity was chosen as the detection system. Biotinylated anti-TSH monoclonal antibody (MAb) and HRP-labeled streptavidin were first synthesized. Then the signal amplification was achieved through the interaction between the biotinylated anti-TSH MAb and the HRP-streptavidin conjugate. The light intensity developed was in proportion to the TSH present in the samples. The assay showed little cross-reactivity with three other glycoprotein hormones (human chorionic gonadotropin (HCG), luteinizing hormone (LH), follicle stimulating hormone (FSH)) due to the high specificity of the antibody. The working range for human thyroid stimulating hormone was 0.1-40 mU L(-1). Both the intra-assay and inter-assay coefficients of variation were less than 10% for the BSAS based chemiluminescent enzyme immunoassay (CLEIA). The proposed assay had a sensitivity of 0.01 mU L(-1) which was 10-fold higher than the HRP-MAb conjugate based TSH immunoassay. Thus the higher sensitivity facilitated the clinical testing for thyroid states. The effects of several reaction parameters, such as incubation time, temperature, and reaction volume of the method, were also studied. This method has been successfully applied to the evaluation of TSH in human serum. Compared with the commercial enzyme chemiluminescent immunoassay, the correlation was satisfied.

Development of a high-throughput, indirect antibody immobilization format chemiluminescence enzyme immunoassay (CLEIA) for the determination of progesterone in human serum.

A high-throughput and simple chemiluminescence (CL) enzyme immunoassay (CLEIA) for the determination of progesterone (P) in human serum was developed, with the highly sensitive 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2'-adamantane) (AMPPD)-alkaline phosphatase (ALP) system as the CL detection system. The results showed that the indirect immobilization of rabbit anti-progesterone polyclonal antibody (RAPA) through secondary antibody exhibited apparent advantages over direct coating in terms of antibody saving and improvement of the coating stability and uniformity. The direct analysis of P in human serum without extraction was realized by using 8-anilino-1-naphthalenesulphonic acid (ANS) to displace P from its binding proteins. The effect of several relevant parameters of the immunoreaction were examined and optimized. Compared with some commercial progesterone kits, the presented CLEIA has higher sensitivity with detection limitation as low as 0.06 ng/mL. The recoveries were 95.9-101%. The coefficient of variation was <8.4% and 9.9% for intra- and inter-assay precision, respectively. This method has been successfully applied to the evaluation of P in human serum.

Development of high-performance magnetic chemiluminescence enzyme immunoassay for alpha-fetoprotein (AFP) in human serum.

BACKGROUND: A high-performance chemiluminescence enzyme immunoassay (CLEIA) for alpha-fetoprotein (AFP), a tumor marker for the diagnosis of hepatocellular carcinoma (HCC), was constructed by using magnetic particles (MPs) as both the immobilization matrix and separation tools. METHODS: A double sandwiched immunocomplex was formed through the reaction among anti-fluorescein isothiocyanate (FITC) antibody coated MPs, FITC-labeled anti-AFP antibody, AFP antigen, and alkaline phosphatase (ALP)-labeled anti-AFP antibody. The subsequent chemiluminescence reaction of ALP with 4-methoxy-4-(3-phosphate-phenyl)-spiro-(1,2-dioxetane-3,2'-adamantane) (AMPPD) gave light intensity that was directly proportional to the amount of analyte present in the samples. The effects of several physicochemical parameters, including the concentration of FITC-labeled anti-AFP antibody, the dilution ratio of ALP-labeled anti-AFP antibody, the volume of MPs and substrate, the immunoreaction time and other relevant variables upon the immunoassay were studied and optimized. RIA and microplate CLEIA were used as comparison methods. RESULTS: The proposed method had a sensitivity of 3.0 ng/ml, low cross reactivities, and an assay time of 1 h. The linear range was 0-1200 ng/ml through using MPs and is useful for samples with extremely high AFP concentrations without dilution while avoiding the hook effect. The intra- and inter-assay precision was <3% and <5%. The present method has been successfully applied to the detection of AFP human serum with recoveries from 90 to 108%, and showed a good correlation with the commercially available AFP RIA kit. CONCLUSIONS: This proposed assay provided apparent advantages over microplate CLEIA and RIA, and facilitated the development of high-throughput screening and automated operation systems in the clinical practice.