Objective To establish a semi-quantitative method for measurement of HCV RNA by use of primers and probe, which was sensitive and designed by ourselves, a new europium fluorescent chelate BHHCT. Methods 44 serum samples of HCV infected patients and 20 samples of the healthy people were collected. HCV RNA in serum sample was extracted by HCV fluorescence PCR diagnostic kit produced by Zhongshan University DAAN Gene Co Ltd, and amplified by RT-PCR in which one PCR primer was pre-labeled with biotin. The amplified products were hybridized with capture probes pre-fixed on the microplate. The biotin in the amplified products was conjugated with europium labeled streptavidin. So europium was linked to the target DNA. Then the fluorescent of europium was measured. Results The linear range of this assay was 10 - 10 copies/ml. Both sensitivity and specificity were 100%. Conclusion Europium labeled RT-PCR assay is a sensitive, specific, fast and non-radioactive contaminant method for the measurement of HCV RNA.

Objective To quantifying the urine human cytomegalovirus(HCMV)DNA from the HCMV infection infants and its corresponding liver function indications,and investigate the relationship between their concentrations.Methods The u-rine samples were collected from HCMV infection infants.HCMV DNA was measured by fluorescence quantitative polymer-ase chain reaction (FQ-PCR).Serum ALT,AST,ALP,GGT,T-Bil and D-Bil liver function indications were detected and the positive rate was analyzed,simultaneously.The correlation between the logarithm urine HCMV DNA (log HCMV DNA) concentration and ALT,AST,ALP,GGT,T-Bil and D-Bil were analyzed by Spearman correlation analysis.Results The dis-tribution range ofurine log HCMV DNA in 444 HCMV infection infants was <2.70~7.90;the positive rate of serum ALT, AST,ALP,GGT,T-Bil and D-Bil were 24.8%,59.0%,95.7%,31.1%,16.7% and 16.3%,respectively.The urine log HC-MV DNA was associated with GGT and the correlation coefficient was 0.099 (P < 0.05),but no associated with ALT, AST,ALP,T-Bil and D-Bil.Conclusion The positive rate of liver function indications will rise in HCMV infection infants, the urine log HCMV DNA was associated with GGT,but not associated with other liver function indications.

Objectives To evaluate accurately hepatocyte injury degree of severe hepatitis patients by quantifying plasma DNA of severe hepatitis patients and study its clinical application in diagnosis of severe hepatitis comparing with ALT.Methods Six milliliters of peripheral blood samples were collected from 185 patients with hepatitis B which are divided into four groups, severe hepatitis with 30 cases, acute hepatitis with 20 cases, chronic B hepatitis with 90 cases, and liver cirrhosis with 45 cases. 100 healthy controls were enrolled. Circulating DNA was extracted from plasma by the BILATEST virus DNA/RNA extraction kit and quantified with a novel duplex real-time PCR assay, respectively.Results Plasma DNA levels of hepatitis B patients were significantly higher than those of healthy controls (104.2 ng/ml vs. 23.4 ng/ml (median),P=0.0000).A significant difference of plasma DNA concentration was found between severe hepatitis and acute hepatitis (P=0.0018), and chronic B hepatitis (P=0.0000), and liver cirrhosis (P=0.0000).The median value of serum ALT of hepatitis B patients was 107.5 U/L, much higher than that of the healthy controls (24.1 U/L,P=0.0000).The levels of serum ALT were significantly different between severe hepatitis and acute hepatitis (P=0.0024), while there was no remarkable difference between severe hepatitis and chronic B hepatitis (P=0.0600), liver cirrhosis (P=1.0000). Moreover, for distinguishing severe hepatitis from liver cirrhosis and chronic B hepatitis, the plasma DNA assay was notably superior to ALT by the analysis of receive operating characteristic (ROC) curves (AUC, 0.95 vs. 0.51,P=0.0000; 0.86 vs. 0.34,P=0.0000).Conclusion The results by measuring plasma DNA of hepatitis B patients with the novel duplex real-time quantitative PCR showed that plasma DNA may be considered as a robust predictive marker for accurately evaluating hepatocyte injury degree of severe hepatitis patients.

Objective To evaluate whether the protein SP70 could be used as a serum biomarker for the diagnosis of non-small cell lung cancer (NSCLC).Methods Polyclonal antibody was prepared by immunizing New Zealand rabbit with SPC-A1 cells.Sandwich ELISA was carried out by using newly-prepared polyclonal antibody(PcMb) coating assay plates,monoclonal antibody (McAb) NJ001 and HRP goat antimouse antibody as primary antibody and labeling antibody respectively.After optimizing the experiment conditions,serum from 175 lung cancer patients [ 80 NSCLC adenocarcinoma,70 NSCLC squamous carcinoma and 25 small cell lung cancer ( SCLC) ],25 benign lung disease ( BLD) patients and 300 healthy controls (HC) were examined.CEA,NSE,CYFRA21-1 were measured by ECLIA for comparison.Results Positive rates of NSCLC adenocarcinoma,NSCLC squamous carcinoma,SCLC and BLD were 68.8％,51.4％,16.0％ and 12.0％ respectively,obviously higher than that of HC (7.3％).NSCLC (adenocarcinoma,Squamous carcinoma) had significantly higher positive rate than SCLC (60.7％ υs 16.0％,x2 =17.23,P＜0.05)and BLD(60.7％ υs 12.0％,x2 =20.41,P ＜0.05).Among 68 NSCLC patients who had definite staging,positive rates at early stage ( Ⅰ/Ⅱ,n=30) reached up to 76.7％.Meanwhile,positive rates of CEA,NSE and CYFRA21-1 (32.7％,18.0％ and 37.3％) were significantly lower than the targeting antigen to McAb NJ001 in NSCLC(60.7％ υs 32.7％,x2 =23.63,P ＜0.05;60.7％υs18.0％,x2 =57.22,P＜0.05;60.7％ υs37.3％,x2=16.34,P＜0.05).Conclusions It showed high positive rates of SP70 in the serum of NSCLC patients,which suggested thai SP70 might be a potential valuable biomarker for the diagnosis of NSCLC.

Objective To investigate the diagnostic value of detection of protein SP70 in differentiating benign and malignant pleural effusion.Methods A case-control study was conducted from July 2011 to February 2012.108 cases of pleural effusion from patients with clinically proven lung cancers and 122 cases of benign pleural effusion were collected.SP70 was detected by Sandwich ELISA,while CEA,CYFRA21-1,NSE were measured by electrochemiluminescence immunoassay for comparison.Meanwhile,protein SP70 on exfoliated cells in pleural effusion was detected by direct immunofluorescence,and was compared with the results of HE staining.The differences between the groups were evaluated by the chisquare test Fisher' s exact test.Results Positive rates of SP70,CEA,CYFRA21-1,NSE were 72.2％,58.3％,52.8％ and 30.6％ in malignant pleural effusion,obviously higher than benign pleural effusio (9.8％,13.1％,23.0％ and 19.7％).The specificity of SP70,CEA,CYFRA21-1,NSE were 90.2％,86.9％,77.0％ and 80.3％,NSCLC had significantly higher positive rate than SCLC(74.3％ ＞0.0％,P =0.02 ＜ 0.05),detection of protein SP70 in malignant pleural effusion had significantly higher coincidence rate than HE staining(72.2％ vs 47.2％,x2 =14.03,P ＜ 0.05).Conclusion Determination of the protein SP70 in pleural effusion and in exfoliated cells,can improve the sensitivity and specificity of the diagnosis of malignant pleural effusion.

Objective: To compare the diagnostic consistency and efficacy of arthroscopy and MRI on the severity of acute lateral ligament complex (LLC) injury, and to find an effective method to diagnose acute LLC injury accurately in early stage. Methods: Data of 37 patients (27 males, 10 females), with acute lateral ligament complex injury who were treated from February 2015 to December 2016 were retrospectively analyzed. The average age was 28.6±6.2 years (range, 18-42 years) and duration from injury to operation was 4.6±3.3 days (range, 0.3-8 days). The results of MRI and ankle arthroscopy for detecting LLC injury were compared, and the consistency between the two examination results was analyzed. Results: Different consistencies were found between the two examination methods according to different parts of LCC injuries. The consistency of injuries (partial tear,complete tear) at the starting point of anterior talofibular ligament and complete tear at the starting point of calcaneofibularliga-ment was ordinary (kappa=0.641, 0.624, 0.680), while the consistency of damage in other parts of LLC was poor (kappa< 0.4). Compared with arthroscopy, MRI has a diagnostic accuracy of 62% (23/37) for partial tear of anterior talofibular ligament, and 62% (23/37) for complete tear. And that of the calcaneofibularligament complete tear was 78% (29/37). Conclusion MRI had low capability in detecting the severity of acute LLC injury accurately and the diagnosing efficacy was insufficient, while ar-throscopic exploration had a high diagnostic value for acute ankle joint injuries.

Objective:To discuss the problems that exist in the operation mechanism of the third -party media-tion mode of medical disputes in China and put forward suggestions for improvement , through comparative study of the third-party mediation mode of domestic and foreign medical disputes .Results:People's mediation mechanism of medical disputes in our country still hadsome problems .For example , mediation talents were insufficient;propa-ganda was inadequate;mediation results were difficult to implement;mediation support funds couldn ' t be guaran-teed, and so on.Conclusion:It is recommended to improve the remuneration of employees and standardize the staff selection system;increase propaganda and raise public awareness;improve the judicial confirmation system and en-hance the guarantee of agreement;save regulation cost and increase multi -financing channels .

BACKGROUND: Circulating levels of cell-free DNA increase in many pathologic conditions. However, notable discrepancies in the quantitative analysis of cell-free DNA from a large number of laboratories have become a considerable pitfall, hampering its clinical application. METHODS: We designed a novel recombinant DNA fragment that could be applied as an internal standard in a newly developed and validated duplex real-time PCR assay for the quantitative analysis of total cell-free plasma DNA, which was tested in 5,442 healthy adults and 200 trauma patients. RESULTS: Compared with two traditional methods, this novel assay showed a lower detection limit of 0.1 ng/mL, lower intra- and inter-assay CVs, and higher accuracy in the recovery test. The median plasma DNA concentration of healthy males (20.3 ng/mL, n=3,092) was significantly higher than that of healthy females (16.1 ng/mL, n=2,350) (Mann-Whitney two-sample rank sum test, P<0.0001). The reference intervals of plasma DNA concentration were 0-45.8 ng/mL and 0-52.5 ng/mL for healthy females and males, respectively. The plasma DNA concentrations of the majority of trauma patients (96%) were higher than the upper normal cutoff values and were closely related to the corresponding injury severity scores (R²=0.916, P<0.0001). CONCLUSIONS: This duplex real-time PCR assay with a new internal standard could eliminate variation and allow for more sensitive, repeatable, accurate, and stable quantitative measurements of plasma DNA, showing promising application in clinical diagnosis.
Adult ; DNA/*blood/standards ; Female ; Healthy Volunteers ; Humans ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction/*methods/standards ; Reference Values ; Wounds and Injuries/blood

To meet the needs of management of medical case information and biospecimen simultaneously, we developed a novel medical case information system integrating with biospecimen management. The database established by MS SQL Server 2000 covered, basic information, clinical diagnosis, imaging diagnosis, pathological diagnosis and clinical treatment of patient; physicochemical property, inventory management and laboratory analysis of biospecimen; users log and data maintenance. The client application developed by Visual C++ 6.0 was used to implement medical case and biospecimen management, which was based on Client/Server model. This system can perform input, browse, inquest, summary of case and related biospecimen information, and can automatically synthesize case-records based on the database. Management of not only a long-term follow-up on individual, but also of grouped cases organized according to the aim of research can be achieved by the system. This system can improve the efficiency and quality of clinical researches while biospecimens are used coordinately. It realizes synthesized and dynamic management of medical case and biospecimen, which may be considered as a new management platform.
Biological Specimen Banks ; Database Management Systems ; Humans ; Medical Records Systems, Computerized ; Medical Records, Problem-Oriented

Objective: To compare the clinical effects of urokinase thrombolytic therapy for optic artery occlusion (OAO) and retinal artery occlusion (RAO) caused by facial microinjection with hyaluronic acid and spontaneous RAO. Methods: From January 2014 to February 2018, 22 eyes of 22 patients with OAO and RAO caused by facial microinjection of hyaluronic acid who received treatment in Xi'an Fourth Hospital were enrolled in this retrospective study (hyaluronic acid group). Twenty-two eyes of 22 patients with spontaneous RAO were selected as the control group. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. FFA was used to measure arm-retinal circulation time (A-Rct) and filling time of retinal artery and its branches (FT). Meanwhile, MRI examination was performed. There were significant differences in age and FT between the two groups (t=14.840, 3.263; P=0.000, 0.003). The differecens of logMAR visual acuity, onset time and A-Rct were not statistically significant between the two groups (t=0.461, 0.107, 1.101; P=0.647, 0.915, 0.277). All patients underwent urokinase thrombolysis after exclusion of thrombolytic therapy. Among the patients in the hyaluronic acid group and control group, there were 6 patients of retrograde ophthalmic thrombolysis via the superior pulchlear artery, 6 patients of retrograde ophthalmic thrombolysis via the internal carotid artery, and 10 patients of intravenous thrombolysis. FFA was reviewed 24 h after treatment, and A-Rct and FT were recorded. Visual acuity was reviewed 30 days after treatment. The occurrence of adverse reactions during and after treatment were observed. The changes of logMAR visual acuity, A-Rct and FT before and after treatment were compared between the two groups using ttest. Results: At 24 h after treatment, the A-Rct and FT of the hyaluronic acid group were 21.05±3.42 s and 5.05±2.52 s, which were significantly shorter than before treatment (t=4.569, 2.730; P=0.000, 0.000); the A-Rct and FT in the control group were 19.55±4.14 s and 2.55±0.91 s, which were significantly shorter than before treatment (t=4.114, 7.601; P=0.000, 0.000). There was no significant difference in A-Rct between the two groups at 24 h after treatment (t=1.311, P=0.197). The FT difference was statistically significant between the two groups at 24 h after treatment (t=4.382, P=0.000). There was no significant difference in the shortening time of A-Rct and FT between the two groups (t=0.330, 0.510; P=0.743, 0.613). At 30 days after treatment, the logMAR visual acuity in the hyaluronic acid group and the control group were 0.62±0.32 and 0.43±0.17, which were significantly higher than those before treatment (t=2.289, 5.169; P=0.029, 0.000). The difference of logMAR visual acuity between the two groups after treatment was statistically significant (t=2.872, P=0.008). The difference in logMAR visual acuity before and after treatment between the two groups was statistically significant (t=2.239, P=0.025). No ocular or systemic adverse reactions occurred during or after treatment in all patients. Conclusions Urokinase thrombolytic therapy for OAO and RAO caused by facial microinjection with hyaluronic acid and spontaneous RAO is safe and effective, with shortening A-Rct, FT and improving visual acuity. However, the improvement of visual acuity after treatment of OAO and RAO caused by facial microinjection with hyaluronic acid is worse than that of spontaneous RAO.