Nomogram for evaluating coronary revascularisation necessity based on multi-parameter coronary CT angiography.

AIM: To build a nomogram model to improve the evaluation of revascularisation necessity using multi-parameter coronary computed tomography (CT) angiography (CCTA). MATERIALS AND METHODS: In this retrospective study, 335 patients who underwent CCTA and required revascularisation within 1 month were selected and allocated to the revascularisation group; 208 patients who did not undergo revascularisation were allocated to the non-revascularisation group. CCTA parameters, including CCTA stenosis, plaque qualitative-quantitative characteristics, and fractional flow reserve derived from CT angiography (CT-FFR), for both groups were analysed and compared. Independent risk factors for evaluating revascularisation were obtained using univariate and multivariable regression analysis, after which multi-parameter models were built. Finally, a nomogram was created with these independent risk factors using the R programming language. RESULTS: Plaque analysis was performed successfully for 543 patients with 1,072 target plaques. The performance of the multi-parameter model (AUC 0.894, p<0.001) was significantly higher than that of models based on stenosis (AUC 0.804, p<0.001), plaque qualitative/quantitative characteristics (AUC 0.754/0.789, p<0.001), or CT-FFR (AUC 0.848, p<0.001) alone, to evaluate the necessity of revascularisation. The independent risk factors were CCTA stenosis (OR 1.004, p=0.04), positive remodelling (OR 2.474, p<0.001), total plaque volume (OR 1.001, p<0.001), non-calcified plaque volume proportion (OR 1.019, p<0.001), and CT-FFR (OR 0.001, p<0.001). Subsequently, a nomogram based on these factors was created. CONCLUSION: The multi-parameter CCTA model improved performance in evaluating revascularisation necessity. The nomogram based on these factors is shows promise in clinical settings.

[Study on mosaicism of SCN1A gene mutation in parents of children with Dravet syndrome].

Objective: To investigate the clinical phenotypes and the mutant allele proportion of parents with SCN1A gene mutation mosaicism of Dravet syndrome (DS) children, thus to provide guidance for family reproduction and prenatal diagnosis. Method: The clinical data and peripheral blood DNA samples of DS patients with a SCN1A gene mutation proved by Sanger sequencing were collected prospectively from February 2005 to November 2016 in Department of Pediatrics, Peking University First Hospital. The same mutation was searched in parents and other available relatives. Parental somatic mosaicism was confirmed and quantified by Ion Torrent Personal Genome Machine (PGM) and Raindrop droplet digital PCR (ddPCR). The families were followed up and prenatal diagnosis was provided. Result: Mosaicisms of SCN1A gene mutation in parents were identified in 5.2% (30 out of 575) DS families. Seventeen were fathers and thirteen were mothers. The mutant allele proportion ranged from 1.7% to 32.9% by PGM and from 0.82% to 34.51% by ddPCR, respectively. In 30 parents with somatic mosaicism, thirteen were asymptomatic, ten had a history of febrile seizures (FS), five with epilepsy, one with febrile seizure plus and one had a history of afebrile seizure. Four families had two children with DS. Three siblings of the probands were confirmed genetically with the same pathogenic mutation. One deceased sister of the proband was assumed to have the same pathogenic mutation because she matched DS diagnosis after medical history review despite no blood sample. Two families received prenatal diagnosis. One second pregnancy was terminated because the fetus inherited the mutation as the mother's wish. Conclusion: Sanger sequencing detects parents of some children with DS are SCN1A mutation mosaics. PGM and ddPCR can be used for accurate quantification of mutant mosaics, which can provide accurate guidance for family genetic counseling.

Cytotoxicity of pilosulin 1, a peptide from the venom of the jumper ant Myrmecia pilosula.

The synthetic peptide pilosulin 1, corresponding to the largest defined allergenic polypeptide found in the venom of the jumper ant Myrmecia pilosula, inhibited the incorporation of [methyl-3H]thymidine into proliferating Epstein-Barr transformed (EBV) B-cells. The LD50 was four-fold lower in concentration than melittin, a cytotoxic peptide found in honey bee venom. Loss of cell viability was assessed by flow cytometry by measuring the proportion of cells that fluoresced in the presence of the fluorescent dye 7-aminoactinomycin D. Examination of proliferating EBV B-cells indicated that the cells lost viability within a few minutes exposure to pilosulin 1. Partial peptides of pilosulin 1 were less efficient in causing loss of cell viability and the results suggest that the 22 N-terminal residues are critical to the cytotoxic activity of pilosulin 1. Normal blood white cells were also labile to pilosulin 1. T- and B-lymphocytes, monocytes and natural killer cells, however, were more labile than granulocytes. Analysis of pilosulin 1 using circular dichroism indicated that, in common with melittin and other Hymenoptera venom toxins, it had the potential to adopt an alpha-helical secondary structure.

Translational motion compensation for coronary angiogram sequences.

A method of compensating for the lag of the video cameras typically used in angiographic systems is presented for use in sequences of digitized X-ray images. The lag effect is reduced by a straightforward weighted subtraction, which has the undesirable side effect of increasing noise. By superimposing several lag-corrected and appropriately shifted images, however, the signal-to-noise ratio can be restored. The algorithm uses the phase-correlation method to measure the two-dimensional shift of a mobile coronary arterial structure. Processing is confined to a rectangular area of interest (AOI), which encloses a feature of clinical significance. The differences of the phases of the Fourier transforms of two frames is computed, combined with an appropriate filter, and inverse Fourier-transformed to produce a phase-correlation image. The vector separation from the origin of image space of the peak of the phase-correlation image is the estimate of the shift of the artery's position in the second frame as compared to the first. The isolation of the AOI from the surrounding image is achieved by the application of a window and correction for any linear trend in the background intensity.

A preliminary study of the inhibiting mechanism of anisodamine on rabbit platelets activated by E. coli endotoxin.

In this study, we observed the endotoxin induced platelet aggregation, 5-HT and beta-g releases, cellular cAMP decrease, and also the changes of morphology and membrane fluidity of platelet. These changes play an important role in septic shock, especially in disseminated intravascular coagulation (DIC) and respiratory distress syndrome (RDS). After anisodamine (654) intervention all the parameters mentioned above were improved to a certain extent. This preliminary study of the inhibiting mechanism of 654 on rabbit platelets activated by ET supports the clinical possibility of using 654 for relieving DIC and RDS.

[Production of monoclonal antibodies against Mycobacterium leprae].

A series of hybridoma cell lines, which secrete monoclonal antibodies (McAbs), were produced by means of fusion between mouse myeloma cells SP2/O and spleen cells from BALB/C mice immunized with whole M. leprae plus unique phenolic glycolipid I(PGL-I) of M. leprae and M. leprae sonicates supernatant fluid (MLSS) as immunogen. Primary identification indicated that H2 cell line can secrete McAb against the epitope of PGL-I; IIIE10 cell line can secrete McAb against PGL-I and MLSS and (5) 24 D6C8 cell line only against whole M. leprae. The uses of these McAbs in serodiagnosis of leprosy, identification of M. leprae, analysis and purification of M. leprae antigens, and key problems in technology for producing McAbs against M. leprae were also discussed.

[Study on rapid serodiagnosis for leprosy].

In this article, we have developed two latex agglutination reaction tests (LAR) with PGL-1 and ND-O-BSA as antigens in 80 cases of leprosy patients (LL = 20, BL = 20, BT = 20, TT = 20), 20 cases of tuberculosis and 25 cases of normal control. These two latex agglutination tests were compared with relevant ELISAs (ND-O-BSA- and PGL-1-ELISA), and analysed by Chi-square (chi 2) test. The results indicated that the sensitivity and specificity of the two LARs (PGL-1- and ND-O-BSA-LAR) are similar to their relevant ELISAs respectively (PGL-1-LAR Vs ND-O-BSA-LAR, chi 2 = 1.785, P greater than 0.10; PGL-1-LAR Vs PGL-1-ELISA, chi 2 = 2.0, P greater than 0.10; ND-O-BSA-LAR Vs ND-O-BSA-ELISA, chi 2 = 0.972, P greater than 0.25), and the increase of positive rate is coincident with clinical classification of leprosy. Since LARs are more simple and rapid than ELISAs, LARs may become more useful tools for immunodiagnosis of leprosy in the field. Also conditions to ensure the stability and reproducibility of LARs are discussed in detail.

[Studies on the serological activity of ND-O-BSA and its application in serodiagnosis of leprosy].

In this article, optimum conditions of ND-O-BSA-ELISA were determined and comparisons made of the serologic activity of 3 different types of antigens (WML, PGI, ND-O-BSA) in leprosy (151) and tuberculosis (20) patients and normal controls (42). The results indicated that: 1) optimum conditions for ND-O-BSA-ELISA were ND-O-BSA and OPD concentration of 0.1 and 400 micrograms/ml respectively with HRP-IgM and test sera dilutions of 1:1000 and 1:200; and 2) serological activity showed highly significant positive correlations between 3 types of antigens (WML vs PGI: r = 0.88, P less than 0.0005; PGI vs ND-O-BSA: r = 0.86, P less than 0.0005), all of which were highly sensitive and specific for leprosy serodiagnosis with positive rates of 100% (PGI) and 97.4% (WML, ND-O-BSA) in leprosy patients. None were positive in normal controls, but crossreactivity of different degree were present in tuberculosis patients. The authors suggest that PGI-ELISA and ND-O-BSA-ELISA be used in combination in the evaluation of clinical therapeutic efficacy and determination of disease stages, as well as in epidemiologic survey of subclinical leprosy infection in "high-risk" areas.

[Detection of specific circulating immune complexes in leprosy patients by mouse monoclonal antibody against phenolic glycolipid-I].

In this article, we first report the development of a new test for detecting specific circulating immune complexes (SCIC) in the sera of leprosy patients: This test was named monoclonal antibody specific binding assay (McAb/SBA). We screened for SCIC (PGLI-IgG, PGLI-IgM) in 200 serum batches from 140 leprosy patients, 20 tuberculosis patients and 40 normal controls, and compared the McAb/SBA with PGL1/ELISA. The results indicated that: 1) McAb/SBA was highly sensitive (90%) and specific (95%). Its Youden's Index was 90. Except for its specificity of 95%, the sensitivity (85%) and Youden's Index (85) of the PGL1/ELISA test were lower than those of McAB/SBA; 2) The positive rate (34/40) in paucibacillary patients using McAb/SBA was higher than that (28/40) in PGL1/ELISA; 3) The increase and decline of MOD values in McAb/SBA were associated with BI. McAb/SBA is a new method for detecting SCIC (PGL1-IgG, PGL1-IgM) in the sera of leprosy patients, and it is more sensitive than PGL1-ELISA.

[Detection of antibodies to sulpholipid IV from mycobacterium tuberculosis in pulmonary tuberculosis patients].

Serum IgG and IgM antibodies against a novel 2,3-diacyl-trehalose-2'-sulphate (SLIV) antigen using ELISA were determined in control (n = 112) and in pulmonary tuberculosis (n = 120) patients. The specificity and sensitivity of SLIV-IgG-ELISA were 96% and 52%, respectively. Those of SLIV-IgM-ELISA were 95% and 33%, respectively. Compared with PPD-IgG-ELISA, SLIV-IgG-ELISA showed that the specificity was about the same but the sensitivity was higher.

[Detection of Mycobacterium tuberculosis using a nested-primer gene amplification assay].

A nested-primer gene amplification assay (NPGAA) was established by using two pairs of primers, an outside pair of primers and an inside pair of primers, sequences of which were from M. tuberculosis(Mtb) gro EL genes. The outside pair of primers should amplify a 576-bp piece of the Mtb gro EL gene that contains sites for the inside pair of primers, which should amplify a 344-bp piece. The results shows NPGAA'S detectable limitation was 1 organism/ml and no amplification products were produced from DNAs of other mycobacterium species tested in this study. For detecting Mtb, the entire NPGAA, from sample preparation to data analysis, can be completed within 6-8 hours. When identifying AFB cultures isolated from lesions of skin diseases, the same positive size patterns were obtained.

Unraveling the Numerous Biosynthetic Products of the Marine Sediment-Derived Fungus, Aspergillus insulicola.

A new tripeptide, pre-sclerotiotide F (3), was isolated from a marine sediment-derived fungus, Aspergillus insulicola, along with five known compounds, one of which was new at the time of isolation, scerotiotide F (4). The absolute configuration elucidation of the new compound was determined using a combination of NMR, HR-ESI-MS, and optical rotation analyses. Cytotoxicities were measured in vitro against selected cancer cells. The effects of pre-sclerotiotide F (3) and sclerotiotide F (4) on LPS-induced NF-κB and iNOS expression were also measured.