Construction and Application of YOLOv3-Based Diatom Identification Model of Scanning Electron Microscope Images / 法医学杂志

OBJECTIVES@#To construct a YOLOv3-based model for diatom identification in scanning electron microscope images, explore the application performance in practical cases and discuss the advantages of this model.@*METHODS@#A total of 25 000 scanning electron microscopy images were collected at 1 500× as an initial image set, and input into the YOLOv3 network to train the identification model after experts' annotation and image processing. Diatom scanning electron microscopy images of lung, liver and kidney tissues taken from 8 drowning cases were identified by this model under the threshold of 0.4, 0.6 and 0.8 respectively, and were also identified by experts manually. The application performance of this model was evaluated through the recognition speed, recall rate and precision rate.@*RESULTS@#The mean average precision of the model in the validation set and test set was 94.8% and 94.3%, respectively, and the average recall rate was 81.2% and 81.5%, respectively. The recognition speed of the model is more than 9 times faster than that of manual recognition. Under the threshold of 0.4, the mean recall rate and precision rate of diatoms in lung tissues were 89.6% and 87.8%, respectively. The overall recall rate in liver and kidney tissues was 100% and the precision rate was less than 5%. As the threshold increased, the recall rate in all tissues decreased and the precision rate increased. The F1 score of the model in lung tissues decreased with the increase of threshold, while the F1 score in liver and kidney tissues with the increase of threshold.@*CONCLUSIONS@#The YOLOv3-based diatom electron microscope images automatic identification model works at a rapid speed and shows high recall rates in all tissues and high precision rates in lung tissues under an appropriate threshold. The identification model greatly reduces the workload of manual recognition, and has a good application prospect.

Construction of a 15-plex Rapid STR Multiplex Amplification System / 法医学杂志

OBJECTIVE@#To establish a 15-plex rapid STR multiplex amplification system.@*METHODS@#Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives.@*RESULTS@#Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure.@*CONCLUSION@#The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.

Perinatal management and outcome of different types of fetal arrhythmia / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the perinatal management and outcome of different types of fetal arrhythmia.</p><p><b>METHODS</b>A retrospective analysis was conducted among the fetuses with arrhythmia identified by M-mode and pulsed Doppler echocardiography in a single institution between October 2003 and December 2010.</p><p><b>RESULTS</b>A total of 130 fetuses were found to have fetal arrhythmia. The most common arrhythmia during pregnancy was extrasystole (n=59), followed by bradycardia (n=23), tachycardia (n=16), atrial flutter (AF, n=3), atrioventricular block (AVB, n=12) and other arrhythmia (n=17). The overall incidence of cardiac anomalies (commonly fetal bradycardia) was 9.2% in these cases. The prognosis of arrhythmia differed significantly between cases of different classifications. The type of fetal arrhythmia (P=0.024), presence of congenital heart defect (CHD, P=0.000) and fetal hydrops (P=0.008) were significant risk factors associated with termination of pregnancy.</p><p><b>CONCLUSION</b>Fetal arrhythmias without CHD or hydrops under close monitoring often have good clinical outcome, while fetal bradycardia is associated with a high mortality rate. CHD and the presence of fetal hydrops are significant risk factors for pregnancy termination.</p>

Effect of xuezhlkang capsule in intervening different Chinese medical syndrome patterns of non-alcoholic fatty liver disease complicated with carotid atherosclerosis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the intervening effects of Xuezhikang Capsule (XZK) on levels of blood lipid and other related indices in patients with different Chinese medical syndrome patterns of non-alcoholic fatty liver disease complicated carotid atherosclerosis (NAFLD-CAS), and to seek out the most appropriate pattern to indicate XZK for making guidance of its utilization.</p><p><b>METHODS</b>Chinese medical syndrome in 74 patients of NAFLD-CAS were classified into 4 patterns, 34 of Pi-deficiency phlegm-dampness pattern (A), 24 of dampness-heat accumulation pattern (B), 12 of phlegm-stasis intertwined pattern (C), and 4 of Gan-Shen yin-deficiency pattern (D). Excepting those of pattern D were excluded due to too small samples, all patients were treated with XZK for 3 months. Blood levels of blood lipids, including triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C), as well as high-sensitivity C-reactive protein (hs-CRP), and tumor necrosis factor alpha (TNF-alpha) were detected and compared before and after treatment.</p><p><b>RESULTS</b>The effective rate of XZK on patients of the three patterns, in A-C order, was 97.06%, 91.67%, 91.67%, respectively, with the optimal overall efficacy showed on pattern A. All the indices detected significantly decreased after treatment in all three patterns (P < 0.01), among them, excepting the difference of TG level between groups showed no significance (P > 0.05), the decrements of others were more significant in pattern A than in other two patterns (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>XZK could reduce the levels of blood lipids, hs-CRP and TNF-alpha in NAFLD-CAS patients, and the Pi-deficiency phlegm-dampness syndrome pattern was the optimal indication of XZK treatment.</p>

Clinical and genetic analysis of a pedigree of Kennedy disease / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To review the clinical and genetic features of a pedigree of Kennedy disease in China.</p><p><b>METHODS</b>The clinical data of patients from a Kennedy disease family were collected. The numbers of trinucleotide CAG repeats in exon 1 of the androgen receptor gene were determined by DNA sequencing and repeat fragment analysis.</p><p><b>RESULTS</b>In the pedigree, 4 patients were identified as Kennedy disease. Clinical manifested with adult-onset, progressive proximal limb muscle weakness and atrophy, gynecomastia, oligospermia were also presented. The number of trinucleotide CAG repeats in exon 1 of the androgen receptor gene was 51 in the proband. The electrophysiological study showed sensory and motor involvement and their serum triglycerides values were elevated significantly.</p><p><b>CONCLUSION</b>Androgen receptors gene testing is the most reliable diagnosing method, the patients suspected as Kennedy disease should have a gene testing of androgen receptors.</p>

Changes of T-cell clonality after induction-cultivation of peripheral T lymphocytes in adoptive immunotherapy for leukemias / 中国实验血液学杂志

This study was purposed to analyze the changes of T-cell clonality after induction of peripheral T lymphocytes by autogenous DC and cytokines in the preparation of adoptive immunotherapy for leukemias. The bone marrow and peripheral blood from 21 leukemia patients at remission stage after treatment and subjected to adoptive immunotherapy were collected. Their DCs and T-cells were stimulated with cytokines and then were mixed to activate T-cells. T-cell receptor beta variable region (TCRBV) families were amplified by RT-PCR, and genescan method and sequencing of the PCR products were used to observe the clonality changes of T-cells before and after the induction and cultivation of T-cells. The flow cytometry was used to identify CD3(+), CD4(+), CD8(+), CD3(+)CD56(+) and CD4(+)CD25str(+)FOXP3(+) cells to disclose the ratio change of cytotoxic T lymphocytes (CTL), helper T-cells, regulatory T-cells and NK T-cells before and after induction and cultivation of T-cells. The results showed that in the 21 patients, most of the 24 TCRBV families presented as oligoclonal distribution on genescan, several families were not expressed, and only a few families remained polyclonal. TCRBV24 was found to be oligoclonal in all of the 21 patients. DNA sequence analysis of TCRBV24 revealed a common motif of VAG in CDR3 in 3 cases and a common motif of GGG in CDR3 in 2 cases. In patient 5, both TCRBV 24 and TCRBV8 contained the same motif of GGG in CDR3. The identical motif in these patients may suggest that these T-cells recognize the same antigen. The peripheral lymphocytes demonstrated recovery of clonal profile on genescan from oligoclonal profile and absence of several families before the induction and cultivation to typical polyclonal profile in all TCRBV families after the induction by DC and cytokines for 13 days. After the induction and cultivation, the number of lymphocytes increased to 3.38 +/- 1.20 times. CD3(+), CD4(+), CD8(+), CD3(+)CD56(+) and CD4(+)CD25str(+)FOX P3(+) cells were 71.1 +/- 11.8%, 26.7 +/- 11.4%, 35.7 +/- 12.9%, 3.1 +/- 1.6% and 0.12 +/- 0.1% respectively before the induction and cultivation, and changed to 95.4 +/- 3.2% (p < 0.01), 27.0 +/- 13.1% (p > 0.01), 55.5 +/- 13.8% (p < 0.01), 9.8 +/- 6.1% (p < 0.01) and 0.22 +/- 0.18% (p < 0.01) respectively after the induction and cultivation. It is concluded that the major action of this induction and cultivation method on T-lymphocytes in vitro is the promotion of CTL and NK T-cell proliferation. In leukemic patients at the remission stage, the TCRBV profile is characterized by the oligoclonal proliferation of T-lymphocytes. Several proliferated clones may have the same motif in CDR3, suggesting the recognition of the same antigen by these lymphocyte clones. Cytokine induction and co-culture with autogenous DCs can stimulate the T-lymphocytes to recover their immunocompetence as manifested by the polyclonal profile and the proliferation of CTL and NK-T cells.

Research siblings identification by Identiffler system and automatic STR genetic / 中国法医学杂志

Objective To evaluate the probability of siblings identification in Identifiler system by using the software of automatic analysis.Methods Using the software of automatic analysis in siblings jdentification.STP genetic typing of 151 pairs of full siblings and 31224 pairs of unrelated individuals from manual simulation were analyzed in 15 STR loci of ldentifiler system.Results Kin probability(W_(FS))of 39.07% full siblings were more than 99.999% while W_(FS) of unrelated individual pairs were 0% .W_(FS) of 60.93% full siblings and 21.3% unrelated individual pairs were all at the range from 99.999% to 1% .W_(FS) of 78.7% unrelated individual pairs 0% full siblings individuals were less than 1% .Therefore,there were notability difference between full siblings and unrelated individual pairs.In addition,testing of 15 STR loci of Identifiler system,it suggested that the pair were siblings when the locus number of the entirely-same is not less than 5 or that of the entirely-different is not more than 1,and that the pair were unrelated individuals when the locus number of the entirely-different is not less than 6 or that of the entirely-8alne is not more than 1.Conclusion The software of automatic analysis and the Identifiler system call be used to siblings identification.

Inhibition effect of diimide G-quadruplex ligand on proliferation of leukemia cells and its molecular mechanisms / 中国实验血液学杂志

This study was aimed to investigate the growth inhibition effect of diimide G-quadruplex ligand on leukemia cells and to explore its molecular mechanisms. K562 leukemia cell lines were treated with various concentrations of the diimide G-quadruples ligand small molecule (0.1 - 10 micromol/L). Trypan blue exclusion assay was used to evaluate the proliferation inhibition. Cell apoptosis was observed using terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL). Telomerase activity was analyzed by telomere repeat amplification protocol. Gene expression was detected by microarray and confirmed by RT-PCR assay. The results showed that diimide small molecule inhibited the proliferation of K562 cells and induced apoptosis of these cells. After treating with diimide G-quadruplex ligand, telomerase activity of K562 cells was reduced and the transcriptional levels of some important genes were changed significantly. These genes were involved in cell apoptosis, cell signaling pathway and other key functions. In conclusion, the diimide G-quadruplex ligand is a small molecule that inhibits the proliferation and induces apoptosis in leukemia cells, and these functions may be related to telomerase inhibition and regulation of some important gene transcription.

Urocortin-postconditioning protects heart from ischemia-reperfusion injury through interfering mitochondria-dependent apoptotic pathway / 重庆医科大学学报

Objective:To investigate the effects of Urocortin-postconditioning on mitochondria-dependentapoptotic pathway.Methods: 24 Wistar rats were randomly divided into 3 groups(n=8):Control group(CON),I/R group(I/R),Urocortin group(UCN).The left ventricular samples in the Control group were collected as pre-ischemia controls through thoracotomy.After isolated heart Langendoff and Neely models were established,the rat hearts in the I/R group and the UCN group were continuously perfused with Krebs-Henseleit (K-H)buffersolution for 30 min,then the rat hearts were arrested by cardioplegia,St.Thomas-2 solution(STH-2)for 30min.After that,the rat hearts in the I/R group were reperfused with K-H buffer solution for 90 min.The rat hearts in the UCN group were reperfused with K-H buffer solution containing 10-8 mol/L Urocortin for 90 min.The releasing of Cyto C protein into cytosol was detected by Western blot.The expression of Caspase-3 protein in cardiomyocyte was detected by immunohistochemical assay.The apoptosis index (AI)of cardiomyocyte was detected by TUNEL.Results:Compared with Control group,the expression ofCaspase-3 and Cyto Cprotein in the I/R group and the UCN group were significantly increased(P

Characteristics of T cell receptors recognizing antiphospholipid syndrome associated antigens / 中国实验血液学杂志

To understand the characteristics of T cell receptors recognizing antiphospholipid syndrome associated antigen, the characteristics of T cells were analyzed using T cell receptor beta variable region (TCRbetaV) gene spectrotyping in a case of antiphospholipid syndrome (APS). The results indicated that in the case of APS there were 2 dominant T cell clones. The TCRbetaVs sequences of the 2 T cell clones showed the TCRbetaVs belonged to 8 and 23 gene families respectively. The peptides of third complementarity-determining regions (CDR3) in the TCRbetaVs were CASSLLVAGGPRAYNEQFFGPG and CASSLAGFGQPQHFGDG. Comparing the motifs in CDR3 with another autoimmune disease, the motif YNEQFFGPG in TCRbetaV8 and motif QHFGDG in TCRbetaV23 were identical with that of idiopathic thrombocytopenic purpura and systemic lupus erythematosus reported before. In conclusion, some T cell clones proliferating in these autoimmune diseases may recognize the same antigens.

Mechanism of CD4+CD25+ and CD8+ regulatory T cells--review / 中国实验血液学杂志

Regulatory T cells (Treg) play a key role in the negative regulation of the immune system, which can inhibit inappropriate self-reactive T cells, as well as limit the range, extent and lasting time of immune responses, so as to inhibit the proliferation of CD4(+) and CD8(+) effector T cells. In view of these properties, there is little doubt about the enormous potential value of Tregs in molecular mechanisms and clinical treatment of human autoimmune disorders, graft-versus-host disease (GVHD) and allergies. In this review, the molecular mechanisms of CD4(+)CD25(+) Treg and CD8(+) Treg were mainly discussed, and challenge with which study on Treg cells is faced and prospects were briefly described.