Objective To evaluate the performance of KRAS gene mutation detection in 2014 external quality assessment ( EQA ) program and discuss the problems in clinical laboratories .Methods The sample panel of 2014 EQA program contained 5 artificial formalin-fixed, paraffin-embedded ( FFPE) samples.The participating laboratories were asked to report their results before the deadline .The scores of EQA and the rate of overall coincidence , false positive and false negative were calculated .Results The EQA program for KRAS testing was set twice a year .In 2014, 58 and 57 valid lab results were submitted respectively.About 79.31%(46/58)and 94.73%(54/57) of the laboratories were correct for all samples. The coincidence rate of positive samples were 93.53% ( 217/232 ) and 96.49% ( 165/171 ) . The coincidence rate for negative ones were 100%(58/58) and 98.25% (112/114).The false-negative ratio was 1.29%( 3/232 ) and 0%.The false-positive ratio was 4.14% ( 12/290 ) and 3.15% ( 9/285 ) . Conclusions The results of 2014 EQA for KRAS gene mutation testing suggested that the performance of laboratories had been improved significantly , however , the false-negative and false-positive results had always been the major problems affecting the accuracy of KRAS mutations testing .Laboratories needed to standardize the testing process and manufacturers should optimize the reagents and the way of interpretation , to guarantee the performance of KRAS gene mutation detection .

This paper reports 32 patients with primary esophageal small cell undifferentiated carcinoma confirmed by histologic examination from April 1972 to January 1989 in our hospital. The 1-, and 3-, 5-year survival rates were 53.1%, 18.8% and 6.3%. 93.7% patients died of local un-control, recurrence or metastasis. The age, sex, tumor size, stenosis, position and the type in X-films did not influence the treatment results. Patients with primary esophageal small cell undifferentiated carcinoma should receive a multimodal treatment instead of surgery alone.

Circulating tumor DNA (ctDNA) has shown great potential in targeted therapy efficacy prediction, monitoring, high-risk population screening, differential diagnosis, minimal residual disease (MRD) monitoring, and prognosis prediction. The detection of specific gene mutations in ctDNA had been included in clinical practice guidelines for certain tumors to predict the drug efficacy and monitor resistance. A small number of approved companion diagnostic reagents have been used in clinical setting. However, the clinical validity of most ctDNA-related biomarkers it still in the research stage. Besides, the establishment and validation of laboratory-developed tests (LDT) are also problems that need to be solved urgently. Therefore, for the moment, the clinical application of ctDNA analysis is like two sides of a coin, with both opportunities and challenges.

Objective To compare the performance of fourth generation HIV antigen/antibody combined detection reagents for HIV early infection samples,international HIV seroconversion panel samples and routine clinical screening samples.Methods Thirty seven early HIV infected samples from the followup gays in Shen Yang between 2009 and 2011,66 seroconversion panel samples from BBI company (U.S.A),NABI company(U.S.A) and NIBSC company(U.K) and 703 routine HIV screening samples in the first hospital of China medical university in October 2010 were collected.All kinds of samples were tested by three diagnostic reagents based on chemiluminescence assay (CLIA),electrochemiluminescence assay (ECLIA) and enzyme-linked immunosorbent assay (ELISA) respectively.The detection sensitivity and specificity of these assays were analyzed.Results For 59 early infected and seroconversion samples,the sensitivities of both ECLIA and CLIA reagent were 96.61％ (95％ CI 91.5％-100.0％),higher than that of the ELISA kit (95％ CI 75.0％-92.9％) (x2 =5.341,P ＜ 0.05),which is 83.93％ ; Comparison among the three reagents for different subtypes of the antibody seroconversion samples showed that ECLIA had the highest sensitivity while CLIA was the lowest ; Detection sensitivity of the three reagents for the P24 antigen is CLIA ＞ ECLIA ＞ ELISA; With detection of 703 clinical routine screening samples,the specificities of three reagents were 100％ (CLIA),99.86％ (ECLIA) and 99.71％ (ELISA) respectively.Conclusions For the sensitivity of the fourth HIV diagnostic reagents CLIA and ECLIA are better than ELISA.The former two reagents are more suitable for identifying earlier HIV infection in clinic.

Malignant tracheoesophageal fistula (MTEF) is pathological communication between the respiratory tracts such as the trachea or bronchia and the esophagus because of malignant tumor dissemination through them.Radiography is an important adjunctive technology in the diagnosis of MTEF,and the location and size of fistula often need the further diagnosis of bronchoscopy and esophagoscopy.The patients are often with an unfavourable prognosis once developed MTEF,and are treated usually with the aim of symptom palliation and life quality improvement.The individual treatment includes esophageal stenting,esophageal exclusion and esophagus bypass,fistula exclusion and repair,radiotherapy and others effective therapy according to the patients condition.These therapies will prolong the life span and improve the life quality of patients.MTEF is not absolute contraindication for chemoradiotherapy.Despite its acute toxicity,this concurrent chemoradiothe rapy protocol appears feasible and effective at closing esophageal malignant fistula,especially in patients in a good general condition and without metastasis.

Objective To label human VEGF targeted bevacizumab (avastin) with 111In and to evaluate the application of 111 In?DTPA?avastin SPECT imaging for tumor diagnosis. Methods DTPA?avastin was prepared by coupling with a bifunctional chelating agent, and then labeled with 111 In to obtain 111 In?DTPA?avastin. The stability and molecular integrity of the labeled radiotracer were studied. Human hepatoma cell ( BEL7404) bearing nude mice tumor model was employed for tumor targeting evaluation. Gamma imaging was acquired after intravenous injection of 18.5 MBq probe. At the end of the observation, animals were sac?rificed for bio?distribution study. Results 111 In?DTPA?avastin tracer was synthesized and purified to a?chieve a radiochemical purity yield above 98% and specific activity up to 185 GBq/nmol. Its stability in 5%BSA was optimal, and the radiochemical purity after incubation for 96 h was over 90%. Gamma imaging re?sults showed that the tracer possessed definite tumor targeting property. Its biodistribution was consistent with that of normal in vivo antibody metabolism while possessing a good tumor?targeting property with a relatively high uptake of (3.8±0.8) %ID/g in tumor tissues 96 h after injection. Conclusions 111 In?DTPA?avastin tracer has good physicochemical properties, in vivo stability and good VEGF targeted binding. 111 In?DTPA?avastin has potential to be a new molecular probe for SPECT imaging.

Objective: To better understand the antagonistic effect of Xiao Long Tong Bi (XLTB), a Chinese herb medicine, on α1-adrenoceptor (α1-AR). Methods: (1) Radio ligand binding assay . Specific　125I-BE2254(2-β(4-hdroxyphenyl)-ethyl amino-methyl-tetralone) binding was measured by incubating membrane of canine cerebral cortex with a single concentration of　125I-BE2254 in the presence of 15 concentrations of XLTB. Half-effectual concentration of inhibition (IC50) and Hill coefficients (nH) were determined by Hill plots. (2) Contractile responses of rat prostate strip in vitro were determined. pKB values for XLTB in competitively inhibiting NE-stimulated contraction of tissues were measured by the method of Ainlakshana. Results: XLTB competitively inhibited binding of　125I-BE2254 to α1-AR in a concentration -dependent manner. IC50 values for XLTB in canine cerebral cortex were (34.0±6.0)　g*L－1, the Hill efficiency value (0.7±0.1) was significantly decreased from unity. Contractile studies showed that XLTB competitively antagonized the NE concentration-response curve with pKB values of (37.0±11.0) g*L－1 or (30.0±8.0) g*L－1 when XLTB concentration was 70 g*L－1 or 170 g*L－1, respectively. The pKB values for XLTB in antagonizing NE-induced contraction of tissues were showed to fit in well with the IC50 values on rat prostate. Conclusion: These results suggest that XLTB appears to be a competitive antagonist for α1-AR.

Objective To evaluate the detectability of HIV antigen-antibody in the window period of acute infection by three HIV antigen-antibody assays.Methods Twenty-two samples of HIV seroconversion serum panels and thirty-seven HIV acute infected plasm samples from our laboratory collected from cohort study of men who have sex with men between 2009 and 2011,were assayed by ECLIA,CLIA and ELISA methods.All assays were evaluated for the ability to detect HIV in the window period,and the sensitivity of each assay for acute samples was analyzed.Chi square test was used for statistical analysis.Results The ability of detecting HIV in the window period of each assay was different.For HIV seroconversion serum panels,the results of ECLIA and CLIA assays were consistent,and the window period was shortened at least 1 to 5 days compared with ELISA assay.For HIV acute samples,all were HIV positive by ECLIA or CLIA assay,but for ELISA assay,94.6％ was positive.For samples before seroconversion,ECLIA and CLIA assay had the same sensitivity (93.5％),which is superior to ELISA assay (71.0％) (x2 =5.14,P ＜0.05).Conclusion The ability of detecting HIV in the window period was different for each assay.The results of ECLIA and CLIA assay are consistent,superior to ELISA assay.

AIM To analyze the mechanism of the adriamycin induced cardiomyopathy. METHODS The patch clamp technique in the whole cell recording was used to study the effect of adriamycin on L type calcium channel current( I Ca L ) in the isolated cardiomyocyte of the guinea pig. RESULTS The current voltage( I U ) curveshowed the bell shape in the control and in 0 1 mmol?L -1 adriamycin. Their peak potentials were about +10 mV. The amlitude of peak calcium current increased from (-0 93?0 05) nA to (-1 31?0 08) nA( P

AIM To investigate the effect of ke tamine on ATP-sensitive K + currents (I K ATP ) in single airway smooth muscle (ASM) cells of asthmatic rat. METHODS Single ASM cells of asthmatic rat were isolated with enzymatic dissociation technique. Effect of ketamine on I K ATP in single ASM cells was studied using the whole-cell configu ration of patch clamp technique. RESULTS Ketamine opened the ATP -sensitive K + channel (K ATP channel) in a dose-dependent manner. When the concentrations of ketamine were 1?10 -7 ,1?10 -6 ,1?10 -5 and 1?10 -4 mol?L -1 , the amplitude values of I K ATP were increased to 63 86?19 33 pA/pF(n=8,P