Clinical laboratory features of Meigs' syndrome: a retrospective study from 2009 to 2018 / 医学前沿

Meigs' syndrome (MS), a rare complication of benign ovarian tumors, is easily misdiagnosed as ovarian cancer (OC). We retrospectively reviewed the clinical laboratory data of patients diagnosed with MS from 2009 to 2018. Serum carbohydrate antigen 125 and HE4 levels were higher in the MS group than in the ovarian thecoma-fibroma (OTF) and healthy control groups (all P < 0.05). However, the serum HE4 levels were lower in the MS group than in the OC group (P < 0.001). A routine blood test showed that the absolute counts and percentages of lymphocytes were significantly lower in the MS group than in the OTF and control groups (all P < 0.05). However, these variables were higher in the MS group than in the OC group (both P < 0.05). The neutrophil-to-lymphocyte ratio (NLR) was also significantly lower, whereas the lymphocyte-to-monocyte ratio was higher in the MS group than in the OC group (both P < 0.05). The NLR, platelet-to-lymphocyte ratio, and systemic immune index were significantly higher in the MS group than in the OTF and control groups (all P < 0.05). The hypoxia-inducible factor-1 mRNA levels were also significantly higher, whereas the glucose transporter 1, lactate dehydrogenase, and enolase 1 mRNA levels were lower in peripheral CD4

Development and Evaluation of a Duplex Real-Time PCR Assay With a Novel Internal Standard for Precise Quantification of Plasma DNA

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.

Correlation between Urine HCMV Virus Load and Liver Function Indications among HCMV-Infected Infants / 现代检验医学杂志

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.

Study on the phenotype and function of CD8+Treg cells induced by ovarian cancer cell / 实用医学杂志

Objective To investigate the phenotype and function of CD8+T cells cultured with SK-OV-3. Methods Transwell coculture experiments were conducted in 24-well plates with inner wells to separate CD8+ T cells and SK-OV-3. After 5 days of culture, CD8+ T cells were washed, and 1 × 106 cells were collected for Foxp3, CD25, CD28, CTLA-4 and GITR mRNA analysis and 2 × 106 cells were collected to detect expression of Foxp3, CD25, CD28, CTLA-4 and GITR in CD8+ T cells by flow cytometry. CD8+T cells that cultured alone or with SK-OV-3 were added at ratios of 1∶0 , 1∶1 , 1∶5 , 1∶10 , and 0∶1 to na?ve CD4+ T cells in 96-well plates. All wells were cultured with the presence of irradiated PBMCs and anti-CD3 antibody. After 72 h, [3H]-thymidine was added for 16 h prior to the determination of proliferation by scintillation counting. Results Compared with CD8+ T cells cultured without SK-OV-3 , the expression of Foxp3 and CTLA-4 was increased and CD28 expression was decreased in CD8+ T cells cultured with SK-OV-3 (both P < 0.001). We found that CD8+T cells cultured with SK-OV-3 significantly suppressed the na?ve CD4+ T cell proliferation induced by the anti-CD3 stimulation in a dose-dependent manner. In contrast, CD8+ T cells cultured without SK-OV-3 did not suppress na?ve CD4+ T cell proliferation. Conclusion Ovarian cancer cell can induce the suppressive CD8+Treg, which is an important link of the immunosuppressive microenvironment in ovarian cancer.

The significance of protein SP70 detection for differentiating benign and malignant pleural effusion / 中华检验医学杂志

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.

A preliminary study on serum protein SP70 as a novel biomarker for the detection of non-small cell lung cancer / 中华检验医学杂志

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.

Research and development of medical case database: a novel medical case information system integrating with biospecimen management / 生物医学工程学杂志

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.

Assessment for hepatocyte injury of severe hepatitis patients by quantitative analysis of plasma DNA / 临床检验杂志

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.

Semi-quantitative measurement of HCV RNA by Europium-labeled gene probe / 中华检验医学杂志

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.

PROTECTIVE EFFECT OF ASTRAGALUS MEMBRANCE ON MICE OF ACUTE TOXOPLASMOSIS / 中国血吸虫病防治杂志

Objective To study the protective effect of Astragalus membrance on mice infected with tachyzoites of Toxoplasma gondii. Methods ICR mice were infected intraperitoneally with 10 5, 10 3, 10 2 tachyzoites of virulent RH strain of Toxoplasma gondii, and the mice were orally treated with Astragalus membrance 0 075 g/d per mouse or Azithromycin [150 mg/(d?kg)] each day starting from day 1 post-infection for 10 days. The survival rate and period were investigated. The parasite loads of livers and lungs of the mice infected with 10 2 tachyzoites were determined by fluorescence PCR methods at 4 day-post-infection (dpi) and 8 dpi. Results When infected with 10 5, 10 3 tachyzoites, treated with Astragalus membrance, the average survived days of the mice were 5 57 days and 6 23 days, and treated with azithromycin were 6 96 days and 8 12 days respectively. The azithromycin group but not the astragli group survived significantly longer than the control(P